Substituted sulfonic acid n- (aminoiminomethyl)phenylalkyl!-azaheterocyclamide compounds

ABSTRACT

The compounds of formula I exhibit useful pharmacological activity and accordingly are incorporated into pharmaceutical compositions and used in the treatment of patients suffering from certain medical disorders. More specifically, they are inhibitors of the activity of Factor Xa. The present invention is directed to compounds of formula I, compositions containing compounds of formula I, and their use, which are for treating a patient suffering from, or subject to, physiological condition which can be ameliorated by the administration of an inhibitor of the activity of Factor Xa.

This application is a continuation-in-part application of PCTUS96/09816, filed Jun. 7, 1996 which designates the United States, whichis a continuation-in-part application of U.S. patent application Ser.No. 08/481,024, filed Jun. 7, 1995 now U.S. Pat. No. 5,612,353.

FIELD OF THE INVENTION

The compounds of formula I exhibit useful pharmacological activity andaccordingly are incorporated into pharmaceutical compositions and usedin the treatment of patients suffering from certain medical disorders.More specifically, they are Factor Xa inhibitors. The present inventionis directed to compounds of formula I, compositions containing compoundsof formula I, and their use, which are for treating a patient sufferingfrom, or subject to, conditions which can be ameliorated by theadministration of an inhibitor of Factor Xa.

Factor Xa is the penultimate enzyme in the coagulation cascade. Bothfree factor Xa and factor Xa assembled in the prothrombinase complex(Factor Xa, Factor Va, calcium and phospholipid) are inhibited bycompounds of formula I. Factor Xa inhibition is obtained by directcomplex formation between the inhibitor and the enzyme and is thereforeindependent of the plasma co-factor antithrombin III. Effective factorXa inhibition is achieved by administering the compounds either by oraladministration, continuous intravenous infusion, bolus intravenousadministration or any other parenteral route such that it achieves thedesired effect of preventing the factor Xa induced formation of thrombinfrom prothrombin.

Anticoagulant therapy is indicated for the treatment and prophylaxis ofa variety of thrombotic conditions of both the venous and arterialvasculature. In the arterial system, abnormal thrombus formation isprimarily associated with arteries of the coronary, cerebral andperipheral vasculature. The diseases associated with thromboticocclusion of these vessels principally include acute myocardialinfarction (AMI), unstable angina, thromboembolism, acute vessel closureassociated with thrombolytic therapy and percutaneous transluminalcoronary angioplasty (PTCA), transient ischemic attacks, stroke,intermittent claudication and bypass grafting of the coronary (CABG) orperipheral arteries. Chronic anticoagulant therapy may also bebeneficial in preventing the vessel luminal narrowing (restenosis) thatoften occurs following PTCA and CABG, and in the maintenance of vascularaccess patency in long-term hemodialysis patients. With respect to thevenous vasculature, pathologic thrombus formation frequently occurs inthe veins of the lower extremities following abdominal, knee and hipsurgery (deep vein thrombosis, DVT). DVT further predisposes the patientto a higher risk of pulmonary thromboembolism. A systemic, disseminatedintravascular coagulopathy (DIC) commonly occurs in both vascularsystems during septic shock, certain viral infections and cancer. Thiscondition is characterized by a rapid consumption of coagulation factorsand their plasma inhibitors resulting in the formation oflife-threatening clots throughout the microvasculature of several organsystems. The indications discussed above include some, but not all, ofthe possible clinical situations where anticoagulant therapy iswarranted. Those experienced in this field are well aware of thecircumstances requiring either acute or chronic prophylacticanticoagulant therapy.

SUMMARY OF THE INVENTION

This invention is directed to the pharmaceutical use of a compound offormula I below for treating a patient suffering from a physiologicaldisorder capable of being modulated by inhibiting the activity of FactorXa, where formula I is as follows: ##STR1## is phenyl or monocyclicheteroaryl;

R is hydrogen, optionally substituted alkyl, optionally substitutedaralkyl, optionally substituted heteroaralkyl, R₆ O(CH₂)_(x) --, R₆ O₂C(CH₂)_(x) --, Y¹ Y² NC(O)(CH₂)_(x) --, or Y¹ Y² N(CH₂)_(x) --;

R₁ is hydrogen, alkyl, hydroxy, alkoxy, Y¹ Y² N--, halogen, --CO₂ R₆,--C(O)NY¹ Y², --(CH₂)_(x) OR₆, --(CH₂)_(x) NY¹ Y², or --CN;

R₂ and R₃ are independently selected from hydrogen, hydroxy, alkoxy, Y¹Y² N--, halogen, --CO₂ R₆, --C(O)NY¹ Y², --(CH₂)_(x) OR₆, --(CH₂)_(x)NY¹ Y², --CN, optionally substituted alkyl, optionally substitutedcycloalkyl, optionally substituted heterocyclyl, optionally substitutedaryl, optionally substituted heteroaryl, optionally substituted aralkyl,optionally substituted heteroaralkyl, optionally substituted aralkenylor optionally substituted heteroaralkenyl, or R₂ and R₃ taken togetherwith the carbon atoms through which they are linked form an optionallysubstituted 5 to 7 membered fused cycloalkyl, optionally substituted 5to 7 membered fused heterocyclyl ring or an optionally substituted 6membered fused aryl, or an optionally substituted 5 to 7 membered fusedheteroaryl ring;

R₄ is hydrogen or optionally substituted lower alkyl, optionallysubstituted aralkyl or optionally substituted heteroaralkyl;

X₁ and X_(1a) are independently selected from hydrogen, optionallysubstituted alkyl, optionally substituted aryl, optionally substitutedaralkyl, optionally substituted heteroaryl or optionally substitutedheteroaralkyl, or X₁ and X_(1a) taken together form oxo;

X₂ and X_(2a) are hydrogen, or taken together form oxo;

X₃ is hydrogen, hydroxy, optionally substituted alkyl, optionallysubstituted aryl, optionally substituted heteroaryl, optionallysubstituted aralkyl or optionally substituted heteroaralkyl, or X₃ andone of X₁ and X_(1a) taken together with the carbon atoms through whichX₃ and one of X₁ and X_(1a) are linked form a 4 to 7 membered cycloalkylor heterocyclyl ring;

X₄ is hydrogen, optionally substituted alkyl or an optionallysubstituted aralkyl;

X₅ and X_(5a) are hydrogen or taken together are ═NR₅ ;

R₅ is hydrogen, R₆ O₂ C--, R₆ O--, cyano, R₆ CO--, optionallysubstituted lower alkyl, nitro or Y¹ Y² N--;

Y¹ and Y² are independently hydrogen, optionally substituted alkyl,optionally substituted aryl, optionally substituted aralkyl oroptionally substituted heteroaralkyl, or Y¹ and Y² taken together withthe N through which Y¹ and Y² are linked form a 4 to 7 memberedheterocyclyl;

X₆ and X_(6a) are independently hydrogen, R₇ R₈ N--, R₉ O--, R₇ R₈NCO--, R₇ R₈ NSO₂ --,

R₇ R₈ NSO₂ N--, R₇ R₈ SO₂ O--, R₉ CO--, --CO₂ R₆, --C(O)NY¹ Y²,--(CH₂)_(x) CO₂ R₆, --(CH₂)_(x) C(O)NY¹ Y², --(CH₂)_(x) OR₆, --(CH₂)_(x)NY¹ Y², halo, cyano or nitro;

R₆ is hydrogen, optionally substituted alkyl, optionally substitutedaralkyl or optionally substituted heteroaralkyl;

R₇ and R₈ are independently hydrogen or optionally substituted loweralkyl, or one of R₇ and R₈ is hydrogen and the other of R₇ and R₈ is R₁₀(O)CCH₂ --or lower acyl;

R₉ is hydrogen, optionally substituted lower alkyl, optionallysubstituted lower acyl or R₁₀ (O)CCH₂ --;

R₁₀ is hydrogen, optionally substituted lower alkyl, optionallysubstituted alkoxy or hydroxy;

A is S or --CH═CH--;

m is 0, 1, 2 or 3;

n is 0, 1, 2 or 3; and

x is 1, 2, 3, 4, or 5, or

a pharmaceutically acceptable salt thereof, an N-oxide thereof, ahydrate thereof or a solvate thereof.

DETAILED DESCRIPTION OF THE INVENTION

As used above, and throughout the description of the invention, thefollowing terms, unless otherwise indicated, shall be understood to havethe following meanings:

Definitions

"Patient" includes both human and other mammals.

"Alkyl" means an aliphatic hydrocarbon group which may be straight orbranched having about 1 to about 20 carbon atoms in the chain. Preferredalkyl groups have 1 to about 12 carbon atoms in the chain. Branchedmeans that one or more lower alkyl groups such as methyl, ethyl orpropyl are attached to a linear alkyl chain. "Lower alkyl" means about 1to about 4 carbon atoms in the chain which may be straight or branched.The alkyl may be substituted with one or more "alkyl group substituents"which may be the same or different, and include halo, cycloalkyl,hydroxy, alkoxy, amino, acylamino, aroylamino, carboxy, alkoxycarbonyl,aralkyloxycarbonyl, heteroaralkyloxycarbonyl or Y¹ Y² NCO--, wherein Y¹and Y² are independently hydrogen, optionally substituted alkyl,optionally substituted aryl, optionally substituted aralkyl oroptionally substituted heteroaralkyl, or Y¹ and Y² taken together withthe N through which Y¹ and Y² are linked form a 4 to 7 memberedheterocyclyl. Exemplary alkyl groups include methyl, trifluoromethyl,cyclopropylmethyl, cyclopentylmethyl, ethyl, n-propyl, i-propyl,n-butyl, t-butyl, n-pentyl, 3-pentyl, methoxyethyl, carboxymethyl,methoxycarbonylethyl, benzyloxycarbonylmethyl,pyridylmethyloxycarbonylmethyl.

"Cycloalkyl" means a non-aromatic mono- or multicyclic ring system ofabout 3 to about 10 carbon atoms. Exemplary monocyclic cycloalkyl ringsinclude cyclopentyl, cyclohexyl and cycloheptyl. The cycloalkyl group isoptionally partially unsaturated or optionally substituted with one ormore cycloalkyl group substituents which may be the same or different,where "cycloalkyl group substituent" includes hydrogen, alkyl, aryl,heteroaryl, aralkyl, heteroaralkyl, hydroxy, hydroxyalkyl, alkoxy,aryloxy, aralkoxy, acyl, aroyl, halo, nitro, cyano, carboxy,alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, acylamino,aroylamino, alkylsulfonyl, arylsulfonyl, heteroarylsulfonyl,alkylsulfinyl, arylsulfinyl, heteroarylsulfinyl, alkylthio, arylthio,heteroarylthio, aralkylthio, heteroaralkylthio, fused cycloalkyl, fusedheterocyclyl, arylazo, heteroarylazo, Y¹ Y² N--, Y¹ Y² NCO-- or Y¹ Y²NSO₂ --, wherein Y¹ and Y² are independently hydrogen, optionallysubstituted alkyl, optionally substituted aryl, optionally substitutedaralkyl or optionally substituted heteroaralkyl, or Y¹ and Y² takentogether with the N through which Y¹ and Y² are linked form a 4 to 7membered heterocyclyl. The aryl group substituents are as definedherein. Exemplary multicyclic cycloalkyl rings include 1-decalin,norbornyl, adamant-(1- or 2-)yl.

"Heterocyclyl" means a non-aromatic monocyclic or multicyclic ringsystem of about 3 to about 10 ring atoms. Preferred rings include about5 to about 6 ring atoms wherein one of the ring atoms is oxygen,nitrogen or sulfur. The heterocyclyl is optionally partially unsaturatedor optionally substituted with one or more heterocyclyl groupsubstituents which may be the same or different, where "heterocyclylgroup substituent" includes hydrogen, alkyl, aryl, heteroaryl, aralkyl,heteroaralkyl, hydroxy, hydroxyalkyl, alkoxy, aryloxy, aralkoxy, acyl,aroyl, halo, nitro, cyano, carboxy, alkoxycarbonyl, aryloxycarbonyl,aralkoxycarbonyl, acylamino, aroylamino, alkylsulfonyl, arylsulfonyl,heteroarylsulfonyl, alkylsulfinyl, arylsulfinyl, heteroarylsulfinyl,alkylthio, arylthio, heteroarylthio, aralkylthio, heteroaralkylthio,fused cycloalkyl, fused heterocyclyl, arylazo, heteroarylazo, Y¹ Y² N--,Y¹ Y² NCO-- or Y¹ Y² NSO₂ --, wherein Y¹ and y² are independentlyhydrogen, optionally substituted alkyl, optionally substituted aryl,optionally substituted aralkyl or optionally substituted heteroaralkyl,or Y¹ and Y² taken together with the N through which Y¹ and Y² arelinked form a 4 to 7 membered heterocyclyl. The heterocyclyl groupsubstituents are as defined herein. Exemplary monocyclic rings includepyrrolidyl, piperidyl, tetrahydrofuranyl, tetrahydrothienyl andtetrahydrothiopyranyl. The thio or nitrogen moiety of the heterocyclylmay also be optionally oxidized to the corresponding N-oxide, S-oxide orS,S-dioxide.

"Aryl" means a 6 to 10 membered aromatic monocyclic or multicyclichydrocarbon ring system. Exemplary aryl include phenyl or naphthyl, orphenyl substituted or naphthyl substituted with one or more aryl groupsubstituents which may be the same or different, where "aryl groupsubstituent" includes hydrogen, alkyl, aryl, heteroaryl, aralkyl,heteroaralkyl, hydroxy, hydroxyalkyl, alkoxy, aryloxy, aralkoxy, acyl,aroyl, halo, nitro, cyano, carboxy, alkoxycarbonyl, aryloxycarbonyl,aralkoxycarbonyl, acylamino, aroylamino, alkylsulfonyl, arylsulfonyl,heteroarylsulfonyl, alkylsulfinyl, arylsulfinyl, heteroarylsulfinyl,alkylthio, arylthio, heteroarylthio, aralkylthio, heteroaralkylthio,fused cycloalkyl, fused heterocyclyl, arylazo, heteroarylazo, Y¹ Y² N--,Y¹ Y² NCO-- or Y¹ Y² NSO₂ --, wherein Y¹ and Y² are independentlyhydrogen, optionally substituted alkyl, optionally substituted aryl,optionally substituted aralkyl or optionally substituted heteroaralkyl,or Y¹ and Y² taken together with the N through which Y¹ and Y² arelinked form a 4 to 7 membered heterocyclyl. The aryl group substituentsare as defined herein. Preferred aryl groups are optionally substitutedphenyl or optionally substituted naphthyl. Preferred aryl groupsubstituents include hydrogen, alkyl, hydroxy, acyl, aryl aroyl,aryloxy, halo, nitro, alkoxy, cyano, alkoxycarbonyl, acylamino,alkylthio, Y¹ Y² N--, Y¹ Y² NCO-- or Y¹ Y² NSO₂ --, where Y¹ and Y² areindependently optionally substituted alkyl, aryl, aralkyl orheteroaralkyl; preferred phenyl group substituents are hydroxy, halogen,alkyl, amino.

"Heteroaryl" means about a 5- to about a 10- membered aromaticmonocyclic or multicyclic hydrocarbon ring system in which one or moreof the carbon atoms in the ring system is/are element(s) other thancarbon, for example nitrogen, oxygen or sulfur. The "heteroaryl" mayalso be substituted by one or more of the above-mentioned "aryl groupsubstituents". Exemplary heteroaryl groups include substitutedpyrazinyl, furanyl, thienyl, pyridyl, pyrimidinyl, isoxazolyl,isothiazolyl, oxazolyl, thiazolyl, pyrazolyl, furazanyl, pyrrolyl,imidazo 2,1-b!thiazolyl, benzofurazanyl, indolyl, azaindolyl,benzimidazolyl, benzothienyl, quinolinyl, imidazolyl and isoquinolinyl.

Where ##STR2## is a monocyclic heteroaryl, then preferred heteroarylsinclude thienyl or pyridyl.

"Aralkyl" means an aryl-alkyl- group in which the aryl and alkyl are aspreviously described. Preferred aralkyls contain a lower alkyl moiety.Exemplary aralkyl groups include benzyl, 2-phenethyl andnaphthlenemethyl.

"Heteroaralkyl" means a heteroaryl-alkyl- group in which the heteroaryland alkyl are as previously described. Preferred heteroaralkyls containa lower alkyl moiety. Exemplary heteroaralkyl groups may containthienylmethyl, pyridylmethyl, imidazolylmethyl and pyrazinylmethyl.

"Aralkenyl" means an aryl-alkenyl- group in which the aryl and alkenylare as previously described. Preferred aralkenyls contain a loweralkenyl moiety. An exemplary aralkenyl group is 2-phenethenyl.

"Heteroaralkenyl" means a heteroaryl-alkenyl- group in which theheteroaryl and alkenyl are as previously described. Preferredheteroaralkenyls contain a lower alkenyl moiety. Exemplaryheteroaralkenyl groups may contain thienylethenyl, pyridylethenyl,imidazolylethenyl and pyrazinylethenyl.

"Hydroxyalkyl" means a HO-alkyl- group in which alkyl is as previouslydefined. Preferred hydroxyalkyls contain lower alkyl. Exemplaryhydroxyalkyl groups include hydroxymethyl and 2-hydroxyethyl.

"Acyl" means an H--CO-- or alkyl-CO-- group in which the alkyl group isas previously described. Preferred acyls contain a lower alkyl.Exemplary acyl groups include formyl, acetyl, propanoyl,2-methylpropanoyl, butanoyl and palmitoyl.

"Aroyl" means an aryl-CO-- group in which the aryl group is aspreviously described. Exemplary groups include benzoyl and 1- and2-naphthoyl,

"Heteroaroyl" means an means an heteroaryl-CO-- group in which theheteroaryl group is as previously described. Exemplary groups includethiophenoyl and pyridinoyl.

"Alkoxy" means an alkyl-O-- group in which the alkyl group is aspreviously described. Exemplary alkoxy groups include methoxy, ethoxy,n-propoxy, i-propoxy, n-butoxy and heptoxy.

"Aryloxy" means an aryl-O-- group in which the aryl group is aspreviously described. Exemplary aryloxy groups include phenoxy andnaphthoxy.

"Aralkyloxy" means an aralkyl-O-- group in which the aralkyl groups isas previously described. Exemplary aralkyloxy groups include benzyloxyand 1- or 2-naphthalenemethoxy.

"Alkylthio" means an alkyl-S-- group in which the alkyl group is aspreviously described. Exemplary alkylthio groups include methylthio,ethylthio, i-propylthio and heptylthio.

"Arylthio" means an aryl-S-- group in which the aryl group is aspreviously described. Exemplary arylthio groups include phenylthio andnaphthylthio.

"Aralkylthio" means an aralkyl-S-- group in which the aralkyl group isas previously described. An exemplary aralkylthio group is benzylthio.

"Y¹ Y² N--" means a substituted or unsubstituted amino group, wherein Y¹and Y² are as previously described. Exemplary groups include amino (H₂N--), methylamino, dimethylamino, diethylamino, pyrrolidine, piperidine,benzylamino, or phenethylamino.

"Alkoxycarbonyl" means an alkyl-O--CO-- group. Exemplary alkoxycarbonylgroups include methoxycarbonyl, ethoxycarbonyl, or t-butyloxycarbonyl.

"Aryloxycarbonyl" means an aryl-O--CO-- group. Exemplary aryloxycarbonylgroups include phenoxycarbonyl and naphthoxycarbonyl.

"Aralkoxycarbonyl" means an aralkyl-O--CO-- group. An exemplaryaralkoxycarbonyl group is benzyloxycarbonyl.

"Y¹ Y² NCO--" means a substituted or unsubstituted carbamoyl group,wherein Y¹ and Y² are as previously described. Exemplary groups arecarbamoyl (H₂ NCO--) and dimethylaminocarbamoyl (Me₂ NCO--).

"Y¹ Y² NSO₂ --" means a substituted or unsubstituted sulfamoyl group,wherein Y¹ and Y² are as previously described. Exemplary groups areaminosulfamoyl (H₂ NSO₂ --) and dimethylaminosulfamoyl (Me₂ NSO₂ --).

"Acylamino" is an acyl-NH-- group wherein acyl is as defined herein.

"Aroylamino" is an aroyl-NH-- group wherein aroyl is as defined herein.

"Alkylsulfonyl" means an alkyl-SO₂ -- group. Preferred groups are thosein which the alkyl group is lower alkyl.

"Alkylsulfinyl" means an alkyl-SO-- group. Preferred groups are those inwhich the alkyl group is lower alkyl.

"Arylsulfonyl" means an aryl-SO₂ -- group.

"Arylsulfinyl" means an aryl-SO-- group.

"Halo" means fluoro, chloro, bromo, or iodo. Preferred are fluoro,chloro or bromo, and more preferred are fluoro or chloro.

Preferred Embodiments

A preferred embodiment of the invention is a method for treating apatient suffering from a physiological disorder capable of beingmodulated by inhibiting an activity of Factor Xa by administering atherapeutically effective amount of a compound of formula I.

Another preferred compound aspect of the invention is the compound offormula I wherein n is 1, and m is 1.

Another preferred compound aspect of the invention is the compound offormula I wherein X₂ and X_(2a) taken together are oxo.

Another preferred compound aspect of the invention is the compound offormula I wherein X₁, X_(1a), X₄ are hydrogen, and X₃ is hydrogen oralkyl.

Another preferred compound aspect of the invention is the compoundformula I wherein X₅ and X_(5a) taken together are ═NR₅ wherein R₅ is R₆O₂ C--.

Another preferred compound aspect of the invention is the compoundformula I wherein X₅ and X_(5a) taken together are ═NR₅ wherein R₅ is--OH.

Another preferred compound aspect of the invention is the compound offormula I wherein X₅ and X_(5a) taken together are ═NR₅ wherein R₅ is H.

Another preferred compound aspect of the invention is the compound offormula I wherein ##STR3## is phenyl and the carbon substituted with X₅,X_(5a) and R₄ HN-- is attached meta relative to the attachment of the--(CH)_(n) N-- moiety.

Another preferred compound aspect of the invention is the compound offormula I wherein ##STR4## is thienyl and the carbon substituted withX₅, X_(5a) and R₄ HN-- is attached in the 2 position relative to thesulfur thereof and the attachment of the --(CH)_(n) N-- moiety in the 4position.

Another preferred compound aspect of the invention is the compound offormula I wherein R is hydrogen, methyl, aralkyl, heteroaralkyl, HO₂CCH₂ --, H₂ NC(O)CH₂ --, or R₆ HNC(O)CH₂ --.

Another preferred compound aspect of the invention is the compound offormula I wherein R₁ is hydrogen, alkyl, or halogen.

Another preferred compound aspect of the invention is the compound offormula I wherein R₂ and R₃ are independently hydrogen, halogen,alkyloxy, amino, aryl, or heteroaryl.

Another preferred compound aspect of the invention is the compound offormula I wherein R₂ and R₃ form an optionally substituted fused aryl oran optionally substituted fused heteroaryl ring wherein the substituentis halogen, alkyl, amino, hydroxy, or alkoxy.

Another preferred compound aspect of the invention is the compound offormula I wherein R₂ and R₃ form an optionally substituted fusedcycloalkyl or an optionally substituted fused heterocyclyl in which theheteroatom is nitrogen wherein the substituent is hydrogen, Y¹ Y² N, oralkyl.

Another preferred compound aspect of the invention is the compound ofclaim 1 wherein ##STR5## is phenyl and one of X₆ and X_(6a) is amino orhydroxy in a para position relative to the ##STR6## moiety.

Another preferred compound aspect of the invention is the compound ofclaim 1 wherein X₆ and X_(6a) are hydrogen.

Another preferred compound aspect of the invention is the compound ofclaim 1 wherein A is S (sulfur).

Another preferred compound aspect of the invention is the compound ofclaim 1 wherein A is --CH═CH-- and R₂ and R₃ form an optionallysubstituted 6 membered heteroaryl ring in which the hetero atom is N andthe substituent is chloro, hydroxy or amino.

Species according to the invention are selected from the groupconsisting of:

3- 3-(S)-(Benzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

3- 3-(S)- (Benzob!thiophene-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

4- 3-(S)-(Benzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-thiophene-2-carboxamidinetrifluoroacetate.

4- 3-(S)- (Benzob!thiophene-2-sulfonyl!-methylamino!-2-oxo-pyrrolidin-1-ylmethyl!-thiophene-2-carboxamidinetrifluoroacetate.

3- 3-(S)-(4-Chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

3- 3-(S)-(6-Chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

3- 3-(S)- (4-Chlorobenzob!thiophene-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

3- 3-(S)- (6-Chlorobenzob!thiophene-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

3- 3-(S)-(5-Chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

3- 3-(S)- (5-Chlorobenzob!thiophene-2sulfonyl)-methyl-amino!-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

3- 3-(S)-(4-Methylbenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

3- 3-(S)-(6-Methylbenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

3- 3-(S)-(5-Methylbenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

3- 3-(S)-(4,6-Dichlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

( 3- 3-(S)-(4,6-Dichlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-phenyl!-iminomethyl)-carbamicacid 2,2,2-trichloroethyl ester.

4-Amino-3- 3-(S)-(4,6-dichlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

4-Hydroxy-3- 3-(S)-(4,6-dichlorobenzob!thiophene-2sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

3- 3-(S)-(6-Fluorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

4-Amino-3- 3-(S)-(6-fluorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

4-Hydroxy-3- 3-(S)-(6-fluorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

4-Amino-3- 3-(S)-(4-chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

4-Hydroxy-3- 3-(S)-(4-chlorobenzob!thiophene-2-sulfamylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

3- 3-(S)-(4-Chloro-thieno3,2-c!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

4-Hydroxy-3- 3-(S)-(4-chloro-thieno3,2-c!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

3- 3-(S)-(5-Chlorothieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

3- 3-(S)-(Thieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinebistrifluoroacetate.

3- 3-(S)-(5-Chlorothieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-hydroxybenzamidinetrifluoroacetate.

3-{3-(S)- (5Chlorothieno3,2-b!pyridine-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl}-benzamidinetrifluoroacetate.

3- 3-(S)-(6-Chlorothieno2,3-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

3- 3-(S)-(Thieno2,3-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinebistrifluoroacetate.

3-{3-(S)- (6Chlorothieno2,3-b!pyridine-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl}-benzamidinetrifluoroacetate.

4-Hydroxy-3- 2-oxo-3-(S)-(5chlorothieno3,2-b!pyridine-2-sulfonylamino)-pyrrolidin-1-ylmethyl}-benzamidinetrifluoroacetate.

4-Hydroxy-3- 2-oxo-3-(S)-(thieno3,2-b!pyridine-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinebistrifluoroacetate.

Hydroxy-3- 2-oxo-3-(S)-(5-chlorothieno3,2-b!pyridine-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-hydroxybenzamidinetrifluoroacetate.

4-Amino-3- 2-oxo-3-(S)-(5chlorothieno3,2-b!pyridine-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

4-Amino-3- 2-oxo-3-(S)-(thieno3,2-b!pyridine-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinebistrifluoroacetate.

4- 3-(S)-(5-Chlorothieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-thiophene-2-carboxamidinetrifluoroacetate.

4- 3-(S)-(5-Chlorothieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-thiophene-2-hydroxycarboxamidinetrifluoroacetate.

4-{3-(S)- (5-Chlorothieno3,2-b!pyridine-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl}-thiophene-2-carboxamidinetrifluoroacetate.

3-{3-(S)-5-(2-Methylsulfanyl-pyrimidin-4-yl)-thiophene-2-sulfonylamino!-2-oxo-pyrrolidin-1-ylmethyl}-benzamidinetrifluoroacetate.

3-{3-(S)-5-(2-Methoxy-pyrimidin-4-yl)-thiophene-2-sulfonylamino!-2-oxo-pyrrolidin-1-ylmethyl}-benzamidinetrifluoroacetate.

3-{3-(S)-5-(2-Amino-pyrimidin-4-yl)-thiophene-2-sulfonylamino!-2-oxo-pyrrolidin-1-ylmethyl}-benzamidineditrifluoroacetate.

3-{3-(S)-(5-(2-Amino-pyrimidin-4-yl)-thiophene-2-sulfonyl!-methylamino)-2-oxo-pyrrolidin-1-ylmethyl}-benzamidineditrifluoroacetate.

3- 3-(S)-(5'-Chloro-2,2'!-bithiophenyl-5-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

4-Amino-3- 3-(S)-benzob!thiophene-2-sulfonylamino)-2-oxopyrrolidin-1-ylmethyl!benzamidinetrifluoroacetate.

4-Amino-3- 6-chlorobenzob!thiophene-2-sulfonylamino)-2-oxopyrrolidin-1-ylmethyl!benzamidinetrifluoroacetate.

4-Amino-3- 6-chlorobenzob!thiophene-2-sulfonylamino)-2-oxopyrrolidin-1-ylmethyl!hydroxybenzamidinetrifluoroacetate.

3-2-Oxo-3-(S)-(5-pyridin-3-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

4-Amino-3-2-oxo-3-(S)-(5-pyridin-3-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidine trifluoroacetate.

4-Hydroxy-3-2-oxo-3-(S)-(5-pyridin-3-yl-thiophene-2-sulfonylamino)-pyrrolidin-1ylmethyl!-benzamidinetrifluoroacetate.

4-Hydroxy-3-2-oxo-3-(S)-(5pyridin-N-oxide-3-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

3-2-Oxo-3-(S)-(5-pyridin-4-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

3-3-(S)-(4-Chloro-thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

3-{3-(S)-5-(5-Chloropyridin-3-yl)-thiophene-2-sulfonylamino!-2-oxopyrrolidin-1-ylmethyl}benzamidinetrifluoroacetate.

3-3-(S)-(4-Chloro-5-pyridin-3-ylthiophene-2-sulfonylamino)-2-oxopyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

4-Hydroxy-3- 3-(S)-(6-chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidetrifluoroacetate.

3-3-(S)-(1-Aminoisoquinoline-6-sulfonylamino)-2-oxopyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

4-Fluoro-3-3-(S)-(5-pyridin-3-yl-thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidetrifluoroacetate.

2-Chloroquinoline-6-sulfonic acid {1-3-(aminoiminomethyl)-benzyl!-2-oxo-pyrrolidin-3-(S)-yl}-amidetrifluoroacetate.

2-Aminoquinoline-6-sulfonic acid {1-3-(aminoiminomethyl)-benzyl!-2-oxo-pyrrolidin-3-(S)-yl}-amideditrifluoroacetate.

Compounds of formula I may be prepared by the application or adaptationof known methods, by which is meant methods used heretofore or describedin the literature.

Compounds of formula I can be prepared by treatment of compounds offormula II ##STR7## in an alcoholic solvent such as ethanol with orwithout a co-solvent such as CH₂ Cl₂ with a hydrogen halide gas such asHCl at about -40° C. to about 100° C. The resulting imidate is thentreated in an alcoholic solvent (such as MeOH) with ammonia gas to givecompounds of formula I in which R₄ ═H and X₅ and X_(5a) taken togetherare ═NR₅ where R₅ is H (amidine). For those compounds in which R₅ ═OH,the imidate is treated with hydroxylamine in an alcoholic solvent. Forcompounds in which R₅ ═C(O)OR₆, the amidine is dissolved in a DMF/CH₂Cl₂ solution followed by treatment with a tertiary amine base such astriethylamine, diisopropylethylamine or N-methylpiperidine and an alkylcarbonate or an alkyl chloroformate at about -20° C. to about 100° C.Compounds of formula II in which R does not equal H can be produced bytreatment of compounds of formula III ##STR8## in an inert organicsolvent, such as THF, DMF, or Et₂ O, with a strong base such as sodiumhydride or potassium carbonate, followed by the addition of anoptionally substituted alkyl halide, optionally substituted aralkylhalide, or an optionally substituted heteroaralkyl halide attemperatures about -78° C. to about 100° C. Compounds of formula III canbe prepared from amines of formula IV ##STR9## by either dissolving theamine hydrochloride in an organic solvent such as CH₂ Cl₂, THF or DMFcontaining an appropriate base such as triethylamine or potassiumcarbonate or by dissolving the amine hydrochloride in an organic basesuch as pyridine followed by addition of a sulfonyl chloride representedby formula V. ##STR10## Sulfonyl chlorides represented by formula V canbe produced by treatment of compounds represented by formula VI##STR11## with a strong base such as n-BuLi at -78° C. followed by theaddition of SO₂ gas and treatment of the lithium heteroaryl sulfonatewith a chlorinating agent such as NCS or SO₂ Cl₂. Compounds of formulaIV in which ##STR12## is a phenyl group substituted as follows;##STR13## can be prepared by the treatment of compounds of formula VIIin which P is a standard protecting moiety such as benzyl, t-butyl,allyl, silylethyl, or trichloroethyl, ##STR14## with a methyl haliderepresented by compounds of formula VIII ##STR15## in an inert organicsolvent such as THF, Et₂ O or DMF in the presence of a strong base suchas NaH, lithium hexamethyldisilylazide or lithium diisopropylamine. Thecarbonate protecting group is then removed by one of the followingappropriate deprotection methods such as acidic, basic, hydrogenolysis,palladium mediated deprotection, zinc promoted deprotection or fluorideanion mediated deprotection. Compounds of formula VIII can be preparedby treating compounds of formula IX ##STR16## with a chlorinating orbrominating agent such as NCS or NBS. When X₆ ═NH₂ and X_(6a) is H, thecorresponding nitro compound is reduced using SnCl₂ and protected as adisubstituted alkyl or aryl or aralkyl imine. The resulting protectedmaterial is brominated using NBS to give a compound represented byformula VIII. When When X₆ ═OH and X_(6a) is H, the hydroxyl group isprotected as silyl, MOM- or MEM- group The resulting compound is thenbrominated using NBS. Compounds of formula IV in which ##STR17## isthienyl group substituted as shown below; ##STR18## can be prepared byreaction of a bromine or iodine containing thiophene carboxaldehyde witha reducing reagent such as sodium borohydride or lithium borohydride.The resulting bromine or iodine containing thiophene methanol is thenconverted to the nitrile using a palladium mediated coupling reactionwith a cyanide source such as Zn(CN)₂. The resultingcyano-thiophene-methanol is then converted to the correspondingmethylchloride or methylbromide using PPh₃ with CCl₄ or CBr₄.

Compounds of formula IV in which X₃ ═H can be obtained by commercialsources or literature sources or by the treatment of a protectedoptionally substituted diamino alkyl carboxylic acid, in which theprotection is at the amino attached to the carbon bearing the carboxylicacid, with peptide coupling reagents such as EDC, DCC or BOP in an inertsolvent such as CH₂ Cl₂, THF or DMF. Compounds in which X₃ ═alkyl can beprepared using methods similar to those described by J. E. Baldwin etal, Tetrahedron 46 (13), p 4733, 1990.

The compounds of the present invention are useful in the form of thefree base or acid or in the form of a pharmaceutically acceptable saltthereof. All forms are within the scope of the invention.

Where the compound of the present invention is substituted with a basicmoiety, acid addition salts are formed and are simply a more convenientform for use; and in practice, use of the salt form inherently amountsto use of the free base form. The acids which can be used to prepare theacid addition salts include preferably those which produce, whencombined with the free base, pharmaceutically acceptable salts, that is,salts whose anions are non-toxic to the patient in pharmaceutical dosesof the salts, so that the beneficial inhibitory effects on the activityof Factor Xa inherent in the free base are not vitiated by side effectsascribable to the anions. Although pharmaceutically addition salts areuseful as sources of are preferred, all acid addition salts are usefulas sources of the free base form even if the particular salt, per se, isdesired only as an intermediate product as, for example, when the saltis formed only for purposes of purification, and identification, or whenit is used as intermediate in preparing a pharmaceutically acceptablesalt by ion exchange procedures. Pharmaceutically acceptable saltswithin the scope of the invention are those derived from the followingacids: mineral acids such as hydrochloric acid, trifluoroacetic acid,sulfuric acid, phosphoric acid and sulfamic acid; and organic acids suchas acetic acid, citric acid, lactic acid, tartaric acid, malonic acid,methanesufonic acid, ethanesulfonic acid, benzenesulfonic acid,p-toluenesulfonic acid, cyclohexylsulfamic acid, quinic acid, and thelike. The corresponding acid addition salts comprise the following:hydrohalides, e.g. hydrochloride and hydrobromide, trifluoroacetate,sulfate, phosphate, nitrate, sulfamate, acetate, citrate, lactate,tartarate, malonate, oxalate, salicylate, propionate, succinate,fumarate, maleate, methylene-bis-B-hydroxynaphthoates, gentisates,mesylates, isethionates and di-p-toluoyltartratesmethanesulfonate,ethanesulfonate, benzenesulfonate, p-toluenesulfonate,cyclohexylsulfamate and quinate, respectively.

According to a further feature of the invention, acid addition salts ofthe compounds of this invention are prepared by reaction of the freebase with the appropriate acid, by the application or adaptation ofknown methods. For example, the acid addition salts of the compounds ofthis invention are prepared either by dissolving the free base inaqueous or aqueous-alcohol solution or other suitable solventscontaining the appropriate acid and isolating the salt by evaporatingthe solution, or by reacting the free base and acid in an organicsolvent, in which case the salt separates directly or can be obtained byconcentration of the solution.

The compounds of this invention may be regenerated from the acidaddition salts by the application or adaptation of known methods. Forexample, parent compounds of the invention can be regenerated from theiracid addition salts by treatment with an alkali, e.g. aqueous sodiumbicarbonate solution or aqueous ammonia solution.

Where the compound of the invention is substituted with an acidicmoiety, base addition salts may be formed and are simply a moreconvenient form for use; and in practice, use of the salt forminherently amounts to use of the free acid form. The bases which can beused to prepare the base addition salts include preferably those whichproduce, when combined with the free acid, pharmaceutically acceptablesalts, that is, salts whose cations are non-toxic to the animal organismin pharmaceutical doses of the salts, so that the beneficial inhibitoryeffects on the activity of Factor Xa inherent in the free acid are notvitiated by side effects ascribable to the cations. Pharmaceuticallyacceptable salts, including for example alkali and alkaline earth metalsalts, within the scope of the invention are those derived from thefollowing bases: sodium hydride, sodium hydroxide, potassium hydroxide,calcium hydroxide, aluminum hydroxide, lithium hydroxide, magnesiumhydroxide, zinc hydroxide, ammonia, ethylenediamine, N-methyl-glucamine,lysine, arginine, ornithine, choline, N,N'-dibenzylethylenediamine,chloroprocaine, diethanolamine, procaine, diethylamine,N-benzylphenethylamine, piperazine, tris(hydroxymethyl)aminomethane,tetramethylammonium hydroxide, and the like.

Metal salts of compounds of the present invention may be obtained bycontacting a hydride, hydroxide, carbonate or similar reactive compoundof the chosen metal in an aqueous or organic solvent with the free acidform of the compound. The aqueous solvent employed may be water or itmay be a mixture of water with an organic solvent, preferably an alcoholsuch as methanol or ethanol, a ketone such as acetone, an aliphaticether such as tetrahydrofuran, or an ester such as ethyl acetate. Suchreactions are normally conducted at ambient temperature but they may, ifdesired, be conducted with heating.

Amine salts of compounds of the present invention may be obtained bycontacting an amine in an aqueous or organic solvent with the free acidform of the compound. Suitable aqueous solvents include water andmixtures of water with alcohols such as methanol or ethanol, ethers suchas tetrahydrofuran, nitriles such as acetonitrile, or ketones such asacetone. Amino acid salts may be similarly prepared.

The base addition salts of the compounds of this invention can beregenerated from the salts by the application or adaptation of knownmethods. For example, parent compounds of the invention can beregenerated from their base addition salts by treatment with an acid,e.g. hydrochloric acid.

Salt forms according to invention also include compounds having aquarternarized nitrogen. The quarternarized salts are formed by methodssuch as by alkylation of a sp³ or sp² hybridized nitrogen in thecompounds.

As will be self-evident to those skilled in the art, some of thecompounds of this invention do not form stable salts. However, acidaddition salts are most likely to be formed by compounds of thisinvention having a nitrogen-containing heteroaryl group and/or whereinthe compounds contain an amino group as a substituent. Preferable acidaddition salts of the compounds of the invention are those wherein thereis not an acid labile group.

As well as being useful in themselves as active compounds, salts ofcompounds of the invention are useful for the purposes of purificationof the compounds, for example by exploitation of the solubilitydifferences between the salts and the parent compounds, side productsand/or starting materials by techniques well known to those skilled inthe art.

Compounds of the present invention may contain asymmetric centers. Theseasymmetric centers may independently be in either the R or Sconfiguration. It will also be apparent to those skilled in the art thatcertain compounds of formula I may exhibit geometrical isomerism.Geometrical isomers include the cis and trans forms of compounds of theinvention having alkenyl or diazenyl (azo) moieties. The presentinvention comprises the individual geometrical isomers and stereoisomersand mixtures thereof.

Such isomers can be separated from their mixtures, by the application oradaptation of known methods, for example chromatographic techniques andrecrystallization techniques, or they are separately prepared from theappropriate isomers of their intermediates, for example by theapplication or adaptation of methods described herein.

The starting materials and intermediates are prepared by the applicationor adaptation of known methods, for example methods as described in theReference Examples or their obvious chemical equivalents.

The present invention is further exemplified but not limited by thefollowing examples which illustrate the preparation of the compoundsaccording to the invention.

In the nuclear magnetic resonance spectra (NMR) the chemical shifts areexpressed in ppm relative to tetramethylsilane. Abbreviations have thefollowing significance: s=singlet; d=doublet; t=triplet; m=multiplet;dd=doublet of doublets; ddd=doublet of doublets of doublets; dt=doubletof triplets, b=broad, bs=broad singlet, q=quartet, AB=AB pattern.

EXAMPLE 1 3- 3-(S)-(Benzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. (2-Oxo-pyrrolidin-3-(S)-yl)-carbamic acid tert-butyl ester

(S)-Boc-Diaminobutyric acid (25 g, 115 mmol), triethylamine (35 g, 344mmol), and 1-hydroxybenzotriazole hydrate (19.3 g, 143 mmol) aredissolved in 300 mL of THF.1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (27.4 g, 143mmol) is added to the solution. The solution is heated at 60° C. over 15min. A white precipitate forms and the solution is kept at 60° C. for 4h. After this time, the solution is filtered and the collected liquid isconcentrated. The crude product is purified by column chromatographyeluting with a gradient of 1% MeOH/CH₂ Cl₂ to 3% MeOH/CH₂ Cl₂ to affordthe title compound (19.6 g, 98 mmol) as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ6.17 (bs, 1H), 5.08 (bs, 1H), 4.12 (m, 1H),3.33 (m, 2H), 2.65 (m, 1H), 2.00 (m, 1H), 1.42 (s, 9H).

B. 1-(3-Cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-carbamic acid tert-butylester

To a solution of 2-oxo-pyrrolidin-3-(S)-yl)-carbamic acid tert-butylester (9.0 g, 45 mmol) and α-bromo-m-toluyl nitrile (9.3 g, 47 mmol) in225 mL of THF/DMF (10:1) at 0° C. is added a 60% mineral oil dispersionof sodium hydride (1.8 g, 46 mmol). The reaction mixture is stirred at0° C. for 0.5 h and then allowed to warm to ambient temperatures. After3 h, the reaction mixture is quenched by the addition of saturated NH₄Cl. The resulting solution is diluted with EtOAc. The layers areseparated. The organic layer is washed with 1N HCl, H₂ O and saturatedNaCl. The organic layer is dried over MgSO₄, filtered and concentrated.The crude product is purified by column chromatography eluting with agradient of 20% EtOAc/hexanes to 40% EtOAc/hexanes to afford the titlecompound (12.7 g, 40 mmol) as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ7.55 (m, 4H), 5.18 (bs, 1H), 4.47 (AB, 2H),4.18 (dd, 1H), 3.21 (m, 2H), 2.60 (m, 1H), 1.42 (s, 9H).

C. 3-(3-(S)-Amino-2-oxo-pyrrolidin-1-ylmethyl)-benzonitrilehydrochloride

To a solution of 1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-carbamicacid tert-butyl ester (9.1 g, 29 mmol) in 150 mL of EtOAc at 0° C. isbubbled HCl gas for 10 min. After this time, the solution is stirred for4 h. The solution is then concentrated to afford the title compound (7.3g, 29 mmol) as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ8.71 (bs, 3H), 7.85 (m, 2H), 7.70 (m, 2H),4.58 (AB, 2H), 4.13 (M, 1H), 3.32 (m, 2H), 2.44 (m, 1H), 2.18 (m, 1H).

D. Benzo b!thiophene-2-sulfonyl chloride

To a solution of thianaphthalene (11.8 g, 88.1 mmol), in 400 mL of THFat -78° C. is added n-BuLi (55 mL of a 1.6M solution in hexanes, 88.1mmol). After 15 min, the solution is added by cannula to a precooled(-78° C.) solution of SO₂ (200 g) in 100 mL of THF. After addition, thesolution is allowed to warm to ambient temperatures. After 0.5 h, thesolution is concentrated. The residue is suspended in hexanes (400 mL)and is cooled to 0° C. To the solution is added SO₂ Cl₂ (12.5 g, 92.5mmol). After stirring for 15 min, the solution is concentrated. Theresidue is dissolved in EtOAc. The organic solution is washed withsaturated NH₄ Cl (aq.), H₂ O and saturated NaCl (aq.). The organic layeris dried over MgSO₄, filtered and concentrated. The crude product isdissolved in CH₂ Cl₂ and filtered through a plug of silica gel. Theorganic solution is then concentrated. The resulting solid is trituratedwith hexanes to give the title compound (12.1 g, 38 mmol) as a whitesolid.

¹ H NMR (CDCl₃, 300 MHz), δ8.16 (s, 1H), 7.97 (m, 2H), 7.57 (m, 2H).

E. Benzo b!thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

To a solution of3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)-benzonitrile hydrochloride(0.36 g, 1.43 mmol) in 15 mL of CH₂ Cl₂ is added benzob!thiophene-2-sulfonyl chloride (0.32 g, 1.38 mmol). To the resultingsolution is added triethylamine (0.29 g, 2.88 mmol). After 16 h, thesolution is diluted with EtOAc. The solution is washed with 1N HCl,saturated NaHCO₃, and saturated NaCl. The organic layer is dried overMgSO₄, filtered and concentrated. The title compound (0.45 g, 1.09 mmol)is obtained as white solid.

¹ H NMR (CDCl₃, 300 MHz) 7.95 (m, 2H), 7.62 (m, 1H), 7.42 (m, 6H), 5.49(bs, 1H), 4.47 (AB, 2H), 3.90 (m, 1H), 3.24 (m, 2H), 2.63 (m, 1H), 2.10(m, 1H).

F. 3- 3-(S)-Benzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

To a solution of benzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-yl!-amide (0.20 g, 0.49 mmol) in20 mL of EtOH:CH₂ Cl₂ (1:1) at 0° C. is bubbled HCl gas for 5 min. Thesolution is then allowed to warm to ambient temperatures. After 16 h,the solution is concentrated. The residue is dissolved in 20 mL of MeOHand NH₃ gas is bubbled through the solution for 5 min. After this time,the solution is heated at 60° C. After 2 h, the solution isconcentrated. The residue is purified by RP-HPLC eluting with a gradientof 10% CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA). Theappropriate fractions are lyophilized to provide the title compound as asolid.

¹ H NMR (DMSO, 300 MHz) δ9.28 (bs, 2H), 9.02 (bs, 2H), 8.62 (m, 2H),8.04 (s, 1H), 8.02 (m, 3H), 7.65 (m, 1H), 7.51 (m, 5H), 4.40 (AB, 2H),4.22 (m, 1H), 3.09 (m, 2H), 2.14 (m, 1H), 1.68 (m, 1H). FAB MS, M+H!⁺=479. Elemental analysis calculated with 1.5 mole of TFA: C=46.07%,H=3.62%, N=9.35%; found: C=46.35%, H=3.83%, N=9.46%.

EXAMPLE 2 3- 3-(S)- (Benzob!thiophene-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. Benzo b!thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-methylamide

To a solution of benzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide (0.25 g, 0.61 mmol)in 3 mL of DMF is added methyl iodide (0.13 g, 0.91 mmol) followed by K₂CO₃ (0.13 g, 0.91 mmol). The solution is stirred at ambient temperaturesfor 6 h. After this time, the solution is diluted with H₂ O and EtOAc.The layers are separated. The organic layer is washed with H₂ O andsaturated NaCl. The organic layer is dried over MgSO₄, filtered andconcentrated. The residue is triturated with Et₂ O to give the titlecompound (0.25 g, 0.59 mmol) as a white solid.

¹ H NMR (CDCl₃, 300 MHz) 7.99 (s, 1H), 7.82 (m, 2H), 7.54 (m, 1H), 7.38(m, 5H), 4.89 (m, 1H), 4.40 (AB, 2H), 3.18 (m, 2H), 2.89 (s, 3H), 2.32(m, 1H), 1.98 (m, 1H).

B. 3- 3-(S)- (Benzob!thiophene-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F usingbenzo b!thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-methylamide as the startingmaterial. The crude product is purified by RP-HPLC eluting with agradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃ CN/HO₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.30 (bs, 2H), 9.05 (bs, 2H), 8.07 (m, 2H),8.00 (m, 1H), 7.65 (m, 1H), 7.53 (m, 5H), 4.93 (m, 1H), 4.42 (AB, 2H),3.16 (m, 2H), 2.81 (s, 3H), 2.08 (m, 1H), 1.89 (m, 1H). FAB MS, M+H!⁺=443. Elemental analysis calculated with 1.5 mole of H₂ O: C=47.34%,H=4.15%, N=9.35%; found: C=47.26%, H=4.15%, N=9.35%.

EXAMPLE 3 4- 3-(S)-(Benzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-thiophene-2-carboxamidinetrifluoroacetate

A. 5-Iodo-thiophene-3-carboxaldehyde

To a solution of thiophene-3-carboxaldehyde (36 g, 321 mmol) in 80 mL ofCCl₄ and 60 mL of H₂ O is added 2.5 mL of conc. H₂ SO₄ in 160 mL ofacetic acid. To the resulting solution is added HIO₃ (14 g, 80 mmol) andI₂ (38 g, 150 mmol). The solution is refluxed for 6 h. After this time,the reaction is cooled to ambient temperatures and 200 mL of CHCl₃ isadded. The layers are separated. The aqueous layer is extracted withCHCl₃. The organic layers are combined and washed with 0.5M Na₂ S₂ O₃,saturated NaHCO₃ and saturated NaCl. The organic layer is dried overMgSO₄, filtered and concentrated. The crude product is purified bycolumn chromatography eluting with gradient of 2% EtOAc/hexanes to 5%EtOAc/hexanes to afford the title compound (20 g, 84 mmol) as a whitesolid.

¹ H NMR (CDCl₃, 300 MHz) δ9.78 (s, 1H), 8.10 (s, 1H), 7.69 (s, 1H).

B. (5-Iodo-thiophene-3-yl)methanol

To a solution of 5-iodo-thiophene-3-carboxaldehyde (42 g, 176 mmol) in800 mL of THF is added NaBH₄ (7.0 g, 185 mmol). After 1 h, the reactionis quenched by the addition of 100 mL of sat. NH₄ Cl. The resultingsolution is diluted with 1 L of EtOAc. The layers are separated. Theorganic layer is washed with H₂ O and saturated NaCl. The organic layeris dried over MgSO₄, filtered and concentrated. The title compound (42g, 175 mmol) is obtained as an oil.

¹ H NMR (CDCl₃, 300 MHz) δ7.18 (s, 2H), 4.63 (s, 2H), 1.92 (bs, 1H).

C. 4-Hydroxymethyl-thiophene-2-carbonitrile

To a solution of (5-iodo-thiophene-3-yl)methanol (42 g, 176 mmol) in 150mL of DMF is added zinc cyanide (12.4 g, 106 mmol) andtetrakis(triphenylphospine)palladium (0) (8.13 g, 7.04 mmol). Thesolution is heated to 80° C. After 6 h, the solution is diluted with 3 Lof EtOAc. The resulting solution is washed with 1N NH₄ OH, H₂ O and sat.NaCl. The organic layer is dried over MgSO₄, filtered and concentrated.The crude product is purified by column chromatography eluting withgradient of 20% EtOAc/hexanes to 30% EtOAc/hexanes to afford the titlecompound (10 g, 72 mmol) as a clear oil.

¹ H NMR (CDCl₃, 300 MHz) δ7.59 (s, 1H), 7.46 (s, 1H), 4.67 (s, 2H), 2.42(bs, 1H).

D. 4-Bromomethyl-thiophene-2-carbonitrile

To a solution of 4-hydroxymethyl-thiophene-2-carbonitrile (10 g, 72mmol), in 360 mL of THF is added triphenyl phosphine (18.3 g, 76 mmol)and carbon tetrabromide (25 g, 76 mmol). After 3 h, the solution isfiltered and concentrated. The crude product is purified by columnchromatography eluting with gradient of 5% EtOAc/hexanes to 10%EtOAc/hexanes to afford the title compound (14 g, 69 mmol) as a whitesolid.

¹ H NMR (CDCl₃, 300 MHz) δ7.62 (s, 1H), 7.49 (s, 1H), 4.42 (s, 2H).

E. 1-(5-Cyanothiophene-3-ylmethyl)-2-oxo-pyrrolidin-3-(S)-yl!-carbamicacid tert-butyl ester

To a solution of (2-oxo-pyrrolidin-3-(S)-yl)-carbamic acid tert-butylester (3.2 g, 16 mmol), prepared as described in EXAMPLE 1, Part A in 80mL of THF:DMF (10:1) at 0° C. is added4-bromomethyl-thiophene-2-carbonitrile (3.23 g, 16 mmol) and sodiumhydride (60% dispersion in oil, 0.67 g, 16.8 mmol). After addition, thesolution was allowed to warm to ambient temperatures. After 2 h, thesolution is quenched by the addition of sat. NH₄ Cl. The solution isdiluted with H₂ O and EtOAc. The layers are separated. The organic layeris washed with H₂ O and sat. NaCl. The organic layer is dried overMgSO₄, filtered and concentrated. The crude product is purified bycolumn chromatography eluting with gradient of 20% EtOAc/CH₂ Cl₂ to 30%EtOAc/CH₂ Cl₂ to afford the title compound (4.0 g, 13.8 mmol) as a whitesolid.

¹ H NMR (CDCl₃, 300 MHz) δ7.51 (s, 1H), 7.45 (s, 1H), 5.12 (bs, 1H),4.42 (AB, 2H), 4.12 (m, 1H), 3.27 (m, 2H), 2.58 (m, 1H), 1.93 (m, 1H),1.42 (s, 9H).

F. 4-(3-(S)-Amino-2-oxo-pyrrolidin-1-ylmethyl)-thiophene-2-carbonitrilehydrochloride

1-(5-Cyanothiophene3-ylmethyl)-2-oxo-pyrrolidin-3-(S)-yl!-carbamic acidtert-butyl ester (4.0 g, 13.8 mmol) is added to a solution of 100 mL ofEtOAc saturated with HCl gas at 0° C. After 3 h, the solution isconcentrated. The title compound (3.3 g, 13.5 mmol) is obtained as awhite solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ8.61 (bs, 3H), 7.96 (s, 1H), 7.82 (s, 1H),5.12 (bs, 1H). 4.42 (AB, 2H), 4.00 (m, 1H), 3.27 (m, 2H), 2.31 (m, 1H),2.03 (m, 1H).

G. Benzo b!thiophene-2-sulfonic acid1-(5-cyanothiophene-3-ylmethyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared as described in EXAMPLE 1, Part E,substituting4-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)-thiophene-2-carbonitrilehydrochloride for3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)-benzonitrile hydrochloride.The crude product is triturated from Et₂ O to give the product as awhite solid.

¹ H NMR (CDCl₃, 300 MHz) δ7.91 (m, 3H), 7.42 (m, 3H), 7.38 (s, 1H), 5.50(bs, 1H), 4.42 (AB, 2H), 3.89 (m, 1H), 3.27 (m, 2H), 2.66 (m, 1H), 2.13(m, 1H).

H. 4- 3-(S)-(Benzo b!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ymethyl!-thiophene-2-carboxamidine trifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F usingbenzo b!thiophene-2-sulfonic acid1-(5cyanothiophene-3-ylmethyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide as thestarting material. The crude product is purified by RP-HPLC eluting witha gradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.20 (bs, 2H), 8.80 (bs, 2H), 8.55 (m, 1H),8.00 (m, 3H), 7.86 (s, 1H), 7.78 (s, 1H), 7.46 (m, 2H), 4.37 (AB, 2H),4.16 (m, 1H), 3.12 (m, 2H), 2.11 (m, 1H), 1.64 (m, 1H). FAB MS, M+H!⁺=435.

EXAMPLE 4 4- 3-(S)-(Benzob!thiophene-2-sulfonyl!-methylamino!-2-oxo-pyrrolidin-1-ylmethyl!-thiophene-2-carboxamidinetrifluoroacetate

A. Benzo b!thiophene-2-sulfonic acid1-(5-cyanothiophene-3-ylmethyl)-2-oxo-pyrrolidin-3-(S)-yl!-methylamide

The title compound is prepared as described in EXAMPLE 2, Part Asubstituting benzo b!thiophene-2-sulfonic acid1-(5-cyanothiophene-3-ylmethyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide forbenzo b!thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide. The crude product istriturated with Et₂ O to give the product as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ7.98 (s, 1H), 7.87 (m, 2H), 7.42 (m, 4H), 7.38(s, 1H), 4.86 (dd, 1H), 4.38 (AB, 2H), 3.22 (m, 2H), 2.89 (m, 3H), 2.36(m, 1H), 2.03 (m, 1H).

B. 4- 3-(S)-(Benzob!thiophene-2-sulfonyl!-methylamino!-2-oxo-pyrrolidin-1-ylmethyl!-thiophene-2-carboxamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F usingbenzo b!thiophene-2-sulfonic acid1-(5-cyanothiophene-3-ylmethyl)-2-oxo-pyrrolidin-3-(S)-yl!-methylamideas the starting material. The crude product is purified by RP-HPLCeluting with a gradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 70% CH₃ CN/H₂ O(0.1% TFA). The appropriate fractions are lyophilized to provide thetitle compound as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.30 (bs, 2H), 9.05 (bs, 2H), 8.07 (m, 2H),8.00 (m, 1H), 7.65 (m, 1H), 7.53 (m, 5H), 4.93 (m, 1H), 4.42 (AB, 2H),3.16 (m, 2H), 2.78 (s, 3H), 2.08 (m, 1H), 1.89 (m, 1H). FAB MS, M+H!⁺=449. Elemental analysis calculated with 1.5 mole of H₂ O: C=42.78%,H=4.10%, N=9.50%; found: C=42.83%, H=3.71%, N=9.27%.

EXAMPLE 5 3- 3-(S)-4-Chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 1-(Chloro-3-(2,2-dimethoxy-ethyl-sulfanyl)-benzene

To a solution of 3-chlorothiophenol (2.4 g, 16.6 mmol) in THF (200 mL)at 0° C. is added bromoacetaldehyde dimethyl acetal (2.8 g, 16.6 mmol).To the solution is added sodium hydride (60% mineral oil dispersion,0.70 g, 17.4 mmol). The reaction is stirred for 16 h, then quenched bythe addition of saturated NH₄ Cl (aq.). The solution is diluted withEtOAc. The organic layer is washed with saturated NaCl (aq.). Theorganic layer is dried over MgSO₄, filtered and concentrated. The crudeproduct is purified by column chromatography eluting with hexanes. Thetitle compound (3.7 g, 15.9 mmol) is obtained as an oil.

¹ H NMR (CDCl₃, 300 MHz) δ7.32 (m, 1H), 7.25 (m, 1H), 7.12 (m, 1H), 4.47(m, 1H), 3.07 (s, 3H), 3.02 (s, 3H).

B. 4-Chloro-benzo b!-thiophene and 6-Chloro-benzo b!-thiophene

A solution containing polyphosphoric acid (8 g) and chlorobenzene (50mL) is heated at reflux. A solution containing1-chloro-3-(2,2-dimethoxy-ethyl-sulfanyl)-benzene (2.7 g, 1.6 mmol) inchlorobenzene (5 mL) is added dropwise to the refluxing polyphosphoricacid solution. After 6 h, the solution is cooled to ambienttemperatures. The solution is diluted with CH₂ Cl₂ and washed with waterand saturated NaCl (aq.). The organic layer is dried over MgSO₄,filtered and concentrated. The crude product is purified by columnchromatography eluting with hexanes to yield the title compounds (2.4 g,9.0 mmol) as a 1:1 isomeric mixture.

¹ H NMR (CDCl₃, 300 MHz) δ7.88 (m, 1H), 7.75 (m, 2H), 7.42 (m, 2H). EIMS, M!⁺ =168, 170, Cl pattern.

C. 4-Chlorobenzo b!thiophene-2-sulfonyl chloride and 6-Chlorobenzob!thiophene-2-sulfonyl chloride

The title compound is prepared as described in EXAMPLE 1, Part Dsubstituting the 4-chlorobenzo b!-thiophene and 6-chlorobenzob!-thiophene mixture for thianaphthalene. The crude product is purifiedby column chromatography eluting with hexanes to yield the titlecompound as well as 4-chlorobenzo b!thiophene-2-sulfonyl chloride aswhite solids.

4-Chlorobenzo b!thiophene-2-sulfonyl chloride

¹ H NMR (CDCl₃, 300 MHz) δ8.32 (m, 1H), 7.81 (m, 1H), 7.53 (m, 2H).

6-Chlorobenzo b!thiophene-2-sulfonyl chloride

¹ H NMR (CDCl₃, 300 MHz) δ8.11 (s, 1H), 7.88 (m, 2H), 7.50 (m, 1H).

D. 4-Chlorobenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared as described in EXAMPLE 1, Part E,substituting 4-chlorobenzo b!thiophene-2-sulfonyl chloride for benzob!thiophene-2-sulfonyl chloride. The crude product is triturated fromEt₂ O to give the product as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.09 (s, 1H), 7.73 (m, 1H), 7.58 (m, 1H), 7.46(m, 5H), 5.76 (bs, 1H), 4.48 (AB, 2H), 3.96 (m, 1H), 3.24 (m, 2H), 2.66(m, 1H), 2.18 (m, 1H).

E. 3- 3-(S)-(4-Chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.

The title compound is prepared as described in EXAMPLE 1, Part F using4-chlorobenzo b!thiophene2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide as the startingmaterial. The crude product is purified by RP-HPLC eluting with agradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 70% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.28 (bs, 2H), 9.07 (bs, 2H), 8.78 (d, 1H),8.08 (d, 1H), 8.01 (s, 1H), 7.66 (m, 1H), 7.54 (m, 5H), 4.41 (AB, 2H),4.29 (m, 1H), 3.13 (m, 2H), 2.18 (m, 1H), 1.70 (m, 1H). FAB MS, M+H!⁺=463, 465, Cl pattern. Elemental analysis calculated with 1.5 mole of H₂O: C=43.75%, H=3.84%, N=9.28%; found: C=43.85%, H=3.50%, N=9.03%.

EXAMPLE 6 3- 3-(S)-(6-Chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 6-Chlorobenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared as described in EXAMPLE 1, Part E,substituting 6-chlorobenzo b!thiophene-2-sulfonyl chloride, prepared asdescribed in EXAMPLE 5, Part C, for benzo b!thiophene-2-sulfonylchloride. The crude product is triturated from Et₂ O to give the productas a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ7.90 (s, 1H), 7.81 (m, 2H), 7.58 (m, 1H), 7.46(m, 5), 5.58 (bs, 1H), 4.46 (AB, 2H), 3.93 (m, 1H), 3.22 (m, 1H), 2.64(m, 1H), 2.12 (m, 1H).

B. 3- 3-(6-Chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using6-chlorobenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-yl!-amide as the startingmaterial. The crude product is purified by RP-HPLC eluting with agradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 70% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.28 (bs, 2H), 9.16 (bs, 2H), 8.68 (d, 1H),8.27 (s, 1H), 8.01 (s, 1H), 7.98 (d, 1H), 7.68 (m, 1H), 7.55 (m, 5H),4.42 (AB, 2H), 4.21 (m, 1H), 3.10 (m, 2H), 2.14 (m, 1H), 1.68 (m, 1H).FAB MS, M+H!⁺ =463, 465, Cl pattern. Elemental analysis calculated with1.5 mole of H₂ O: C=43.75%, H=3.84%, N=9.28%; found: C=43.85%, H=3.50%,N=9.03%.

EXAMPLE 7 3- 3-(S)- (4-Chlorobenzob!thiophene-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 4-Chlorobenzo b!thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-methylamide

The title compound is prepared as described in EXAMPLE 2, Part Asubstituting 4-chlorobenzo b!thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide for benzob!thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide. The crude product istriturated with Et₂ O to give the product as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.12 (s, 1H), 7.73 (d, 1H), 7.58 (m, 1H), 7.42(m, 5H), 4.90 (t, 1H), 4.41 (AB, 2H), 3.20 (m, 2H), 2.91 (s, 3H), 2.40(m, 1H), 2.04 (m, 1H).

B. 3- 3-(S)- (4-Chlorobenzob!thiophene-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using4-chlorobenzo b!thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-methylamide as the startingmaterial. The crude product is purified by RP-HPLC eluting with agradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 70% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.28 (bs, 2H), 9.10 (bs, 2H), 8.10 (d, 1H),8.03 (s, 1H), 7.58 (m, 1H), 7.52 (m, 5H), 4.97 (t, 1H), 4.40 (AB, 2H),3.15 (m, 2H), 2.77 (s, 3H), 2.15 (m, 1H), 1.94 (m, 1H). FAB MS, M+H!⁺=477, 479, Cl pattern. Elemental analysis calculated with 1.5 mole of H₂O; C=44.70%, H=4.08%, N=9.06%; found: C=44.67%, H=3.66%, N=8.91%.

EXAMPLE 8 3- 3-(S)- (6-Chlorobenzob!thiophene-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 6-Chlorobenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-methylamide

The title compound is prepared as described in EXAMPLE 2, Part Asubstituting 6-chlorobenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide for benzob!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-yl!-amide. The crude product istriturated with Et₂ O to give the product as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ7.95 (s, 1H), 7.84 (m, 2H), 7.58 (m, 1H), 7.43(m, 4H), 4.90 (dd, 1H), 4.41 (AB, 2H), 3.20 (m, 2H), 2.89 (s, 3H), 2.38(m, 1H), 2.04 (m, 1H).

B. 3- 3-(S)- (6Chlorobenzob!thiophene-2sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using6-chlorobenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-methyl-mide as thestarting material. The crude product is purified by RP-HPLC eluting witha gradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 70% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.28 (bs, 2H), 8.98 (bs, 2H), 8.29 (d, 1H),8.12 (s, 1H), 7.99 (d, 1H), 7.65 (m, 1H), 7.52 (m, 4H), 4.89 (t, 1H),4.40 (AB, 2H), 3.13 (m, 2H), 2.74 (s, 3H), 2.08 (m, 1H), 1.90 (m, 1H).FAB MS, M+H!⁺ =477, 479, Cl pattern. Elemental analysis calculated with2.0 mole of H₂ O: C=44.05%, H=4.18%, N=8.93%; found: C=44.13%, H=3.55%,N=8.61%.

EXAMPLE 9 3- 3-(S)-(5-Chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 1-Chloro-4-(2,2-dimethoxy-ethyl-sulfanyl)-benzene

The title compound is prepared as described in EXAMPLE 5, Part Asubstituting 4-chlorothiophenol for 3-chlorothiophenol. The crudeproduct is purified by column chromatography eluting with a gradient of4% Et₂ O/hexanes to 10% Et₂ O/hexanes to afford the title compound as anoil.

¹ H NMR (CDCl₃, 300 MHz) δ7.28 (m, 2H), 7.20 (m, 2H), 4.50 (t, 1H), 3.06(s, 3H), 3.03 (s, 3H). EI MS, M!⁺ =232, 234, Cl pattern.

B. 5-Chlorobenzo b!thiophene

The title compound is prepared as described in EXAMPLE 5, Part Bsubstituting 1-chloro-4-(2,2-dimethoxy-ethyl-sulfanyl)-benzene for1-chloro-3-(2,2-dimethoxy-ethyl-sulfanyl)-benzene. The crude product ispurified by column chromatography eluting with hexanes to afford thetitle compound as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ7.78 (m, 2H), 7.50 (d, 1H), 7.28 (m, 2H). EIMS, M!⁺ =168, 170, Cl pattern.

C. 5-Chlorobenzo b!thiophene-2-sulfonyl chloride

The title compound is prepared as described in EXAMPLE 1, Part Dsubstituting 5-chlorobenzo b!-thiophene for thianaphthalene. The crudeproduct is purified by column chromatography eluting with hexanes toyield the title compound as white solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.05 (s, 1H), 7.98 (s, 1H), 7.82 (d, 1H), 7.53(d, 2H).

D. 5-Chlorobenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-yl!-amide

The title compound is prepared as described in EXAMPLE 1, Part E,substituting 5-chlorobenzo b!thiophene-2-sulfonyl chloride for benzob!thiophene-2-sulfonyl chloride. The crude product is purified by columnchromatography eluting with a gradient of 10% EtOAc/CH₂ Cl₂ to 20%EtOAc/CH₂ Cl₂ to give the product as a white solid.

¹ H NMR (DMSO, 300 MHz) δ8.70 (d, 1H), 8.08 (m, 2H), 7.98 (s, 1H), 7.70(m, 1H), 7.60 (m, 5H), 7.51 (m, 5H), 4.36 (AB, 2H), 4.24 (m, 1H), 3.08(m, 2H), 2.12 (m, 1H), 1.61 (m, 1H). FAB MS, M+H!⁺ =446, 448, Clpattern.

E. 3- 3-(S)-(5Chlorobenzob!thiophene-2sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using5-chlorobenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-methyl-amide as thestarting material. The crude product is purified by RP-HPLC eluting witha gradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.28 (bs, 2H), 8.98 (bs, 2H), 8.69 (d, 1H),8.10 (m, 2H), 7.99 (s, 1H), 7.68 (m, 1H), 7.48 (m, 4H), 4.40 (AB, 2H),4.21 (m, 1H), 3.10 (m, 2H), 2.13 (m, 1H), 1.68 (m, 1H). FAB MS, M+H!⁺=463, 465, Cl pattern. Elemental analysis calculated with 2.0 mole of H₂O C=43.10%, H=3.95%, N=9.14%; found: C=43.33%, H=3.45%, N=8.90%.

EXAMPLE 10 3- 3-(S)- (5-Chlorobenzob!thiophene-2-sulfonyl-methyl-amino!-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 5-Chlorobenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-methylamide

The title compound is prepared as described in EXAMPLE 2, Part Asubstituting 5-chlorobenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide for benzob!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide. The crude productis triturated with Et₂ O to give the product as a white solid.

¹ H NMR (DMSO-d₆ , 300 MHz) δ8.12 (s, 1H), 8.04 (m, 1H), 7.72 (m, 1H),7.64 (s, 1H), 7.50 (m, 4H), 4.91 (m, 1H), 4.37 (AB, 2H), 3.14 (m, 2H),2.76 (s, 3H), 2.06 (m, 1H), 1.88 (m, 1H). FAB MS, M+H!⁺ =460, 462, Clpattern.

B. 3- 3-(S)-(5Chlorobenzob!thiophene-2sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using5-chlorobenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-methylamide as thestarting material. The crude product is purified by RP-HPLC eluting witha gradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 70% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.26 (bs, 2H), 8.95 (bs, 2H), 8.05 (m, 3H),7.62 (m, 1H), 7.48 (m, 4H), 4.90 (t, 1H), 4.39 (AB, 2H), 3.14 (m, 2H),2.77 (s, 3H), 2.08 (m, 1H), 1.89 (m, 1H). FAB MS, M+H!⁺ =477, 479, Clpattern. Elemental analysis calculated with 1.25 mole of H₂ O; C=45.10%,H=4.03%, N=9.15%; found: C=44.97%, H=3.94%, N=8.91%.

EXAMPLE 11 3- 3-(S)-(4-Methylbenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 3-Methyl-1-(2.2-dimethoxy-ethyl-sulfanyl)-benzene

The title compound is prepared as described in EXAMPLE 5, Part Asubstituting 3-methylthiophenol for 3-chlorothiophenol. The crudeproduct is purified by column chromatography eluting with a gradient ofhexanes to 10% Et₂ O/hexanes to afford the title compound as an oil.

¹ H NMR (CDCl₃, 300 MHz) δ7.20 (m, 3H), 7.02 (m, 1H), 4.50 (m, 1H), 3.07(s, 3H), 3.02 (s, 3H), 2.32 (s, 3H). EI MS, M!⁺ =212.

B. 4-Methylbenzo b!thiophene and 6-Methylbenzo b!thiophene

The title compound is prepared as described in EXAMPLE 5, Part Bsubstituting 3-methyl-1-(2,2-dimethoxy-ethyl-sulfanyl)-benzene for1-chloro-3-(2,2-dimethoxy-ethyl-sulfanyl)-benzene. The crude productsare purified by column chromatography eluting with hexanes to afford thetitle compounds as an inseparable (2:1) mixture as an oil. EI MS, M!⁺=148.

C. 4-Methylbenzo b!thiophene-2-sulfonyl chloride and 6-Methylbenzob!thiophene-2-sulfonyl chloride

The title compound is prepared as described in EXAMPLE 1, Part Dsubstituting 4-methylbenzo b!-thiophene and 6-methylbenzo b!-thiophenemixture for thianaphthalene. The crude product is purified by columnchromatography eluting with hexanes to yield 4-methylbenzob!thiophene-2-sulfonyl chloride as well as 6-methylbenzob!thiophene-2-sulfonyl chloride as white solids. 4-Methylbenzob!thiophene-2-sulfonyl chloride

¹ H NMR (CDCl₃, 300 MHz) δ8.22 (s, 1H), 7.78 (d, 1H), 7.49 (t, 2H), 7.32(d, 1H), 2.65 (s, 3H).

6-Methylbenzo b!thiophene-2-sulfonyl chloride

¹ H NMR (CDCl₃, 300 MHz) δ8.09 (s, 1H), 7.83 (d, 2H), 7.70 (s, 1H), 7.35(d, 1H), 2.50 (s, 3H).

D. 4-Methylbenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared as described in EXAMPLE 1, Part E,substituting 4-methylbenzo b!thiophene-2-sulfonyl chloride for benzob!thiophene-2-sulfonyl chloride. The crude product is triturated withEt₂ O to give the product as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.02 (s, 1H), 7.68 (d, 1H), 7.57 (m, 1H), 7.42(m, 4H), 7.23 (m, 1H), 5.49 (bs, 1 H), 4.46 (AB, 2H), 3.90 (m, 1H), 3.23(m, 2H), 2.68 (m, 1H), 2.62 (s, 3H), 2.16 (m, 1H).

E. 3- 3-(S)-(4-Methylbenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate The title compound is prepared as described in EXAMPLE1, Part F using 4-methylbenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide as the startingmaterial. The crude product is purified by RP-HPLC eluting with agradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.26 (bs, 2H), 9.00 (bs, 2H), 8.58 (m, 1H),7.97 (m, 2H), 7.79 (s, 1H), 7.60 (m, 1H), 7.49 (m, 3H), 7.31 (m, 1H),4.40 (AB, 2H), 4.22 (m, 1H), 3.12 (m, 2H), 2.40 (s, 3H), 2.10 (m, 1H),1.66 (m, 1H). FAB MS, M+H!⁺ =443.

Elemental analysis calculated with 2.0 mole of H₂ O; C=46.61%, H=4.59%,N=9.45%; found: C=46.75%, H=4.14%, N=9.38%.

EXAMPLE 12 3- 3-(S)-(6-Methylbenzob!thiophene-2-sulfonylamino-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 6-Methylbenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared as described in EXAMPLE 1, Part E,substituting 6-methylbenzo b!thiophene-2-sulfonyl chloride, prepared asdescribed in EXAMPLE 11, Part C, for benzo b!thiophene-2-sulfonylchloride. The crude product is purified by column chromatography elutingwith a gradient of 10% EtOAc/CH₂ Cl₂ to 15% EtOAc/CH₂ Cl₂ to afford thetitle compound as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ7.89 (s, 1H), 7.79 (d, 1H), 7.67 (s, 1H), 7.58(m, 1H), 7.43 (m, 3H), 7.30 (m, 1H), 5.50 (bs, 1H), 4.2 (AB, 2H), 3.88(m, 1H), 3.21 (m, 2H), 2.64 (m, 1H ), 2.49 (s, 3H), 2.12 (m, 1H).

B. 3- 3-(S)-(6-Methylbenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using5-methylbenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide as the startingmaterial. The crude product is purified by RP-HPLC eluting with agradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.23 (bs, 2H), 9.00 (bs, 2H), 8.56 (d, 1H),7.96 (s, 1H) 7.84 (m, 2H), 7.67 (m, 1H), 7.53 (m, 3H), 7.28 (d, 1H),4.42 (AB, 2H), 4.19 (m, 1H), 3.12 (m, 2H), 2.40 (s, 3H), 2.12 (m, 1H),1.64 (m, 1H). FAB MS, M+H!⁺ =443.

Elemental analysis calculated with 0.50 mole of H₂ O: C=48.84%, H=4.28%,N=9.91%; found: C=48.89%, H=4.05%, N=9.73%.

EXAMPLE 13 3- 3-(S)-5-Methylbenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 4-Methyl-1-(2.2-dimethoxy-ethyl-sulfanyl)-benzene

The title compound is prepared as described in EXAMPLE 5, Part Asubstituting 4-methylthiophenol for 3-chlorothiophenol. The crudeproduct is purified by column chromatography eluting with hexanes toafford the title compound as an oil.

¹ H NMR (CDCl₃, 300 MHz) δ7.28 (m, 2H), 7.10 (m, 2H), 4.49 (t, 1H), 3.06(s, 3H), 3.03 (s, 3H), 2.28 (s, 3H).

B. 5-Methylbenzo b!thiophene

The title compound is prepared as described in EXAMPLE 5, Part Bsubstituting 4-methyl-1-(2,2-dimethoxy-ethyl-sulfanyl)-benzene for1-chloro-3-(2,2-dimethoxy-ethyl-sulfanyl)-benzene. The crude product ispurified by column chromatography eluting with hexanes to afford thetitle compound as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ7.78 (d, 1H), 7.62 (s, 1H), 7.39 (d, 1H), 7.23(d, 1H), 7.17 (d, 1H), 2.50 (s, 3H).

C. 5-Methylbenzo b!thiophene2-sulfonyl chloride

The title compound is prepared as described in EXAMPLE 1, Part Dsubstituting 5-chlorobenzo b!thiophene for thianaphthalene. The crudeproduct is purified by column chromatography eluting with hexanes toyield the title compound as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.08 (s, 1H), 7.78 (m, 2H), 7.39 (d, 1H), 2.51(s, 3H).

D. 5-Methylbenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared as described in EXAMPLE 1, Part E,substituting 5-methylbenzo b!thiophene-2-sulfonyl chloride for benzob!thiophene-2-sulfonyl chloride. The crude product is triturated withEt₂ O to give the product as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ7.86 (s, 1H), 7.73 (m, 2H), 7.69 (s, 1H), 7.60(m, 1H), 7.42 (m, 2H), 7.30 (s, 1H), 5.52 (bs, 1H), 4.43 (AB, 2H), 3.91(m, 1H), 3.20 (m, 2H), 2.64 (m, 1H), 2.48 (s, 3H), 1.61 (m, 1H).

E. 3- 3-(S)-(5-Methylbenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using5-methylbenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide as the startingmaterial. The crude product is purified by RP-HPLC eluting with agradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.26 (bs, 2H), 9.00 (bs, 2H), 8.58 (m, 1H),7.97 (m, 2H), 7.79 (s, 1H), 7.60 (m, 1H), 7.49 (m, 3H), 7.31 (m, 1H),4.40 (AB, 2H), 4.22 (m, 1H), 3.12 (m, 2H), 2.40 (s, 3H), 2.10 (m, 1H),1.66 (m, 1H). FAB MS, M+H!⁺ =443.

Elemental analysis calculated with 0.75 mole of H₂ O C=48.46%, H=4.33%,N=9.83%; found: C=48.41%, H=3.98%, N=9.43%.

EXAMPLE 14 3- 3-(4,6-Dichlorobenzob!thiophene-2-sulfonylamino-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 1,3-Dichloro-5-(2,2-dimethoxy-ethyl-sulfanyl)-benzene

The title compound is prepared as described in EXAMPLE 5, Part Asubstituting 3,5-dichlorothiophenol for 3-chlorothiophenol. The crudeproduct is purified by column chromatography eluting with hexanes toafford the title compound as an oil.

¹ H NMR (CDCl₃, 300 MHz) δ7.19 (s, 2H), 7.12 (s, 1H), 4.51 (m, 1H), 3.13(s, 3H), 3.09 (s, 3H).

B. 4,6-Dichlorobenzo b!thiophene

The title compound is prepared as described in EXAMPLE 5, Part Bsubstituting 1,3-dichloro-5-(2,2-dimethoxy-ethyl-sulfanyl)-benzene for1-chloro-3-(2,2-dimethoxy-ethyl-sulfanyl)-benzene. The crude product ispurified by column chromatography eluting with hexanes to afford thetitle compound as a white solid. EI MS, M!⁺ =202.

C. 4,6-Dichlorobenzo b!thiophene-2-sulfonyl chloride

The title compound is prepared as described in EXAMPLE 1, Part Dsubstituting the 4,6-dichlorobenzo b!thiophene for thianaphthalene. Thecrude product is purified by column chromatography eluting with hexanesto yield the title compound as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.27 (s, 1H), 7.79 (s, 1H), 7.24 (s, 1H).

D. 4,6-Dichlorobenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared as described in EXAMPLE 1, Part E,substituting 4,6-dichlorobenzo b!thiophene-2-sulfonyl chloride for benzob!thiophene-2-sulfonyl chloride. The crude product is triturated withEt₂ O to give the product as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.01 (s, 1H), 7.70 (s, 1H), 7.58 (m, 1H), 7.42(m, 4H), 5.66 (bs, 1H), 4.41 (AB, 2H), 3.95 (m, 1H), 3.22 (m, 2H), 2.62(m, 1H), 2.12 (m, 1H).

E. 3- 3-(S)-(4,6-Dichlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using4,6-dichlorobenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide as the startingmaterial. The crude product is purified by RP-HPLC eluting with agradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 80% CH₂ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.27 (bs, 2H), 9.18 (bs, 2H), 8.82 (m, 1H),8.24 (d, 1H), 7.98 (d, 1H), 7.72 (m, 1H), 7.63 (m, 1H), 7.52 (m, 3H),4.39 (AB, 2H), 4.28 (m, 1H), 3.12 (m, 2H), 2.13 (m, 1H), 1.68 (m, 1H).FAB MS, M+H!⁺ =497, 499, Cl pattern.

Elemental analysis calculated with 1.33 mole of H₂ O: C=43.22%, H=3.44%,N=8.82%; found: C=43.10%, H=3.18%, N=8.47%.

EXAMPLE 15 ( 3- 3-(S)-(4,6-Dichlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-phenyl!-iminomethyl)-carbamicacid 2,2,2-trichloroethyl ester

A. ( 3- 3-(S)-(4,6-Dichlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-phenyl!-iminomethyl)-carbamicacid 2,2.2-trichloroethyl ester

To a solution of 3- 3-(S)-(4,6-dichlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate (0.25 g, 0.40 mmol), prepared as in EXAMPLE 14, Part E,in 4 mL of CH₂ Cl₂ :DMF (10:1) is added N-methyl piperidine (0.12 g, 1.2mmol) followed by trichloroethyl chloroformate (0.93 g, 0.44 mmol). Thesolution is stirred for 16 h. After this time, the solution is dilutedwith EtOAc. The organic layer is washed with 1N HCl, H₂ O, saturatedNaCl. The organic layer is dried over MgSO₄, filtered and concentrated.The residue is purified by column chromatography eluting with a gradientof 1% MeOH/CH₂ Cl₂ to 3% MeOH/CH₂ Cl₂ to afford the title compound (0.20g, 0.30 mmol) as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.23 (bs, 2H), 8.82 (d, 1H), 8.28 (s, 1H),8.00 (s, 1H), 7.84 (d, 1H), 7.76 (s, 1H), 7.72 (s, 1H), 7.40 (m, 2H),4.83 (s, 2H), 4.38 (AB, 2H), 4.27 (m, 1H), 3.09 (m, 2H), 2.13 (m, 1H),1.65 (m, 1H). FAB MS, M+H!⁺ =671, 673, 675, 5-Cl pattern.

EXAMPLE 16 4Amino-3- 3-(S)-4,6-dichlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 4-Amino-3-methyl benzonitrile

To a solution of 3-methyl-4-nitrobenzonitrile (2.0 g, 12.3 mmol) in 100mL of EtOH is added SnCl₂ (13.9 g, 61.7 mmol). The resulting solution isheated at reflux. After 2 h, the solution is cooled to ambienttemperatures. The solution is poured into 150 mL of ice water. The pH ofthe solution is adjusted to >7 with a solution of saturated NaHCO₃. Thesolution is diluted with EtOAc and the resulting mixture is filteredthrough Celite. The filtered solution is separated. The organic layer isdried over MgSO₄, filtered and concentrated to give the title compound(1.57 g, 8.7 mmol) as an off-white solid.

¹ H NMR (CDCl₃, 300 MHz) δ7.30 (m, 2H), 6.63 (d, 1H), 4.10 (bs, 2H),2.15 (m, 2H). EI MS, M!⁺ =132.

B. 4-(Benzhydrylidene-amino)-3-methyl-benzonitrile

To a solution of 4-amino-3-methyl benzonitrile (1.20 g, 9.08 mmol) in 75mL of toluene is added benzophenone (1.74 g, 9.53 mmol) andp-toluenesulfonic acid (0.43 g, 2.1 mmol). The reaction vessel is fittedwith a Dean-Stark trap and the solution is heated at reflux. After 24 h,the solution is cooled to ambient temperatures. The solution isconcentrated. The crude material is purified by column chromatographyeluting with a gradient of 3% EtOAc/hexanes to 10% EtOAc/hexanes. Thetitle compound (2.43 g, 8.2 mmol) is obtained as an oil.

¹ H NMR (CDCl₃, 300 MHz) δ7.80 (m, 2H), 7.40 (m, 6H), 7.30 (s, 1H), 7.15(d, 1h), 7.05 (bs, 2H), 6.50 (d, 1H), 2.20 (s, 3H). EI MS, M!⁺ =296.

C. 4-(Benzhydrylidene-amino)-3-bromomethyl-benzonitrile

To a solution of 4-(benzhydrylidene-amino)-3-methyl-benzonitrile (1.36g, 4.27 mmol) in 40 mL of CCl₄ is added N-bromosuccinimide (0.84 g, 4.7mmol) and benzoyl peroxide (0.22 g, 0.64 mmol). The solution is heatedto reflux for 16 h. The solution is cooled to ambient temperatures. Thesolution is diluted with CH₂ Cl₂. The solution is washed with 1N NaOHand saturated NaCl. The organic layer is dried over MgSO₄, filtered andconcentrated. The crude material is purified by column chromatographyeluting with a gradient of 5% EtOAc/hexanes to 10% EtOAc/hexanes. Thetitle compound (0.91 g, 2.43 mmol) is obtained as an oil.

¹ H NMR (CDCl₃, 300 MHz) δ7.80 (m, 2H), 7.60 (d, 1H), 7.35 (m, 8H), 7.15(dd, 1H), 6.35 (d, 1H), 4.55 (s, 2H). EI MS, M!⁺ =374.

D. {1-2-(Benzhydrylidene-amino)-5-cyano-benzyl!-2-oxo-pyrrolidin-3-(S)-yl}-carbamicacid tert-butyl ester

The title compound is prepared as described in EXAMPLE 1, Part Bsubstituting 4-(benzhydrylidene-amino)-3-bromomethyl-benzonitrile forα-bromo-m-toluyl nitrile. The crude material is purified by columnchromatography eluting with a gradient of 30% EtOAc/hexanes to 40%EtOAc/hexanes. The title compound is obtained as a yellow solid.

¹ H NMR (CDCl₃, 300 MHz) δ7.70 (bs, 2H), 7.40 (s, 1H), 7.38 (bs, 6H),7.30 (d, 1H), 7.15 (bs, 2H), 6.48 (d, 1H), 5.00 (d, 1H, 4.45 (AB, 2H),4.15 (m, 1H), 3.30 (m, 2H), 2.61 (m, 1 H), 1.90 (m, 1H), 1.45 (s, 9H).

E. 4-Amino- 3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)!-benzonitriledihydrochloride

The title compound is prepared as described in EXAMPLE 1, Part C. Thetitle compound is obtained as white solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.68 (bs, 3H), 7.38 (s, 1H), 6.76 (d, 1H),5.68 (bs, 3H), 4.25 (AB, 2H), 4.07 (m, 1H), 3.29 (m, 2H), 2.38 (m, 1H),1.98 (m, 1H).

F. 4,6-Dichlorobenzo b!thiophene-2-sulfonic acid1-(2-amino-5-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared as described in EXAMPLE 1, Part E,substituting 4,6-dichlorobenzo b!thiophene-2-sulfonyl chloride for benzob!thiophene-2-sulfonyl chloride and substituting 4-amino-3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)!-benzonitriledihydrochloride for3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)-benzonitrile hydrochloride.The crude product is triturated with Et₂ O to give the product as awhite solid.

¹ H NMR (CDCl₃ +CD₃ OD, 300 MHz) δ7.97 (s, 1H), 7.68 (s, 1H), 7.40 (s,1H), 7.27 (d, 1H), 7.21 (d, 1H), 6.55 (bs, 1H), 4.20 (AB, 2H), 4.00 (t,1H), 3.14 (m, 2H), 2.45 (m, 1H), 1.90 (m, 1H).

G. 4Amino-3- 3-(S)-(4,6dichlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using4,6-dichlorobenzo b!thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-yl!-amide as the starting material.The crude product is purified by RP-HPLC eluting with a gradient of 10%CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA). The appropriatefractions are lyophilized to provide the title compound as a whitesolid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.27 (bs, 2H), 9.18 (bs, 2H), 8.82 (m, 1H),8.24 (d, 1H), 7.98 (d, 1H), 7.72 (m, 1H), 7.63 (m, 1H), 7.52 (m, 3H),4.39 (AB, 2H), 4.28 (m, 1H), 3.12 (m, 2H), 2.13 (m, 1H), 1.68 (m, 1H).FAB MS, M+H!⁺ =497, 499, Cl pattern.

Elemental analysis calculated with 1.33 mole of H₂ O: C=43.22%, H=3.44%,N=8.82%; found: C=43.10%, H=3.18%, N=8.47%.

EXAMPLE 17 4-Hydroxy-3- 3-(S)-(4,6-dichlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 2-Hydroxy-5-iodo-benzaldehyde

To a solution of salicylaldehyde (10 g, 82 mmol) in 50 mL of CH₂ Cl₂ isadded 22 mL of a 1M ICl solution in CH₂ Cl₂. The solution is stirred for14 h. After this time, a saturated solution of sodium sulfite is addeduntil the color is discharged. The solution is diluted with CH₂ Cl₂. Thelayers are separated. The organic layer is washed with water. Theorganic layer is dried over MgSO₄, filtered and concentrated. Theresulting crude material is recrystallized from cyclohexane to give thetitle compound (7.2 g, 32 mmol) as white solid.

¹ H NMR (CDCl₃, 300 MHz) δ10.91 (s, 1H), 9.82 (s, 1H), 7.84 (d, 1H),7.75 (dd, 1H), 6.75 (d, 1H). EI MS, M!⁺ =248.

B. 5-Iodo-2-(2-methoxy-ethoxymethoxy)-benzyl alcohol

To a solution of sodium hydride (1.2 g of a 60% mineral oil dispersion,52 mmol) in 25 mL of THF at 0° C., is added2-hydroxy-5-iodo-benzaldehyde (7.0 g, 28 mmol). To the resultingsolution is added 2-methoxy-ethoxymethoxy chloride (3.4 mL, 30 mmol) and4 mL of 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone. The solutionis allowed to warm to ambient temperatures. After 45 min., the solutionis cooled to -15° C. and 6 mL of a 2M solution of sodium borohydride inTHF is added. The solution is stirred for 10 min. After this time, 24 mLof a 2M HCl solution in water is added. The resulting solution isdiluted with ether, washed with water and saturated NaCl. The organiclayer is dried over MgSO₄, filtered and concentrated. The resultingcrude material is purified by column chromatography eluting with 40%EtOAc/hexanes to give the title compound (7.6 g, 22.5 mmol) as a whitesolid.

¹ H NMR (CDCl₃, 300 MHz) δ7.64 (d, 1H), 7.52 (dd, 1H), 6.90 (d, 1H),5.30 (s, 2H), 4.62 (d, 2H), 3.82 (m, 2H), 3.54 (m, 2H), 3.35 (s, 3H),2.53 (t, 1H). EI MS, M!⁺ =338.

C. 5-Iodo-2-(2-methoxy-ethoxymethoxy)-benzyl bromide

To a solution of 5-iodo-2-(2-methoxy-ethoxymethoxy)-benzyl alcohol (7.5g, 22 mmol) in 60 mL of THF at 15° C. is added triphenylphospine (6.35g, 24 mmol) followed by N-bromosuccinamide (4.3 g, 24 mmol). Thesolution is stirred for 5 min. The solution is allowed to warm toambient temperature. After 20 min, the solution is concentrated. Thecrude product is purified by column chromatography eluting withEtOAc:CH₂ Cl₂ :hexane (3:1:6).

¹ H NMR (CDCl₃, 300 MHz) δ7.60 (d, 1H), 7.50 (dd, 1H), 6.91 (s, 1H),5.30 (s, 2H), 4.43 (dd, 1 h), 3.84 (m, 2H), 3.53 (m, 2H), 3.35 (s, 3H).EI MS, M!⁺ =400.

D.3-(S)-(tert-Butoxy-carbonyl-amino)-1-(5-iodo-2-(2methoxy-ethoxymethoxy)-benzyl)-pyrrolidin-2-one

The title compound is prepared as described in EXAMPLE 1, Part Bsubstituting 5-iodo-2-(2-methoxy-ethoxymethoxy)-benzyl bromide forα-bromo-m-toluyl nitrile. The crude material is purified by columnchromatography eluting with EtOAc:CH₂ Cl₂ :hexane (3:1:1).

¹ H NMR (CDCl₃, 300 MHz) δ7.53 (dd, 1H), 7.45 (d, 1H), 6.93 (d, 1H),5.26 (s, 2H), 5.20 (bs, 1H), 4.50 (d, 1H), 4.45 (d, 1H), 4.19 (m, 1H),3.80 (m, 2H), 3.53 (m, 2H), 3.37 (s, 3H), 3.22 (m, 2H), 2.62 (m, 1H),1.84 (m, 1H), 1.46 (s, 9H). Ion spray MS, M+H!⁺ =521.

E.3-(S)-(tert-Butoxy-carbonyl-amino)-1-(5-cyano-2-(2-methoxy-ethoxymethoxy)-benzyl)-pyrrolidin-2-one

The title compound is prepared as described in EXAMPLE 3, Part C,substituting3-(S)-(tert-butoxy-carbonyl-amino)-1-(5-iodo-2-(2-methoxy-ethoxymethoxy)-benzyl)-pyrrolidin-2-onefor (5-iodo-thiophene-3-yl)methanol. The crude material is purified bycolumn chromatography eluting with EtOAc:CH₂ Cl₂ :hexane (3:1:1).

¹ H NMR (CDCl₃, 300 MHz) δ7.54 (dd, 1H), 7.45 (d, 1H), 7.20 (d, 1H),5.35 (s, 2H), 5.20 (bs, 1H), 4.50 (d, 1H),1.45 (s, 9H), 4.45 (d, 1H),4.15 (m, 1H), 3.77 (m, 2H), 3.51 (m, 2H), 3.35 (s, 3H), 3.24 (m, 2H),2.60 (m, 1H), 1.90 (m, 1H). EI MS, M!⁺ =420.

F. 3-(S)- (3-Amino-2-oxo-pyrrolidin-1-yl)-methyl!-4-hydroxy-benzonitrilehydrochloride

The title compound is prepared as described in Example 1, Part C,substituting3-(S)-(tert-butoxy-carbonyl-amino)-1-(5-cyano-2-(2-methoxy-ethoxymethoxy)-benzyl)-pyrrolidin-2-onefor 1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-carbamic acidtert-butyl ester. The title compound is obtained as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ8.50 (bs, 3H), 7.50 (dd, 1H), 7.40 (d, 1H),7.00 (d, 1H), 4.26 (d, 2H), 4.00 (m, 1H), 3.25 (m, 2H), 2.30 (m, 1H),1.90 (m, 1H). EI MS, M!⁺ =231.

G. 4,6-Dichlorobenzo b!thiophene-2-sulfonic acid1-(2-hydroxy-5-cyano-benzyl)-2-oxo-pyrrolidin-3-yl!-amide

To a solution of 3-(3-amino-2-oxo-pyrrolidin-1-yl)-methyl!-4-hydroxy-benzonitrilehydrochloride (0.20 g, 0.75 mmol) in 3 mL of pyridine at 0° C. is added4,6-dichlorobenzo b!thiophene-2-sulfonyl chloride. The solution isallowed to warm to ambient temperatures and is stirred for 6 h. Afterthis time, the solution is concentrated. The residue is dissolved in CH₂Cl₂. The organic solution is washed with 1N HCl and saturated NaCl. Thecrude product is triturated with Et₂ O to give the title compound (0.22g, 0.44 mmol) as a white solid.

¹ H NMR (CDCl₃ +DMSO-d₆, 300 MHz) δ7.97 (s, 1H), 7.73 (m, 1H), 7.67 (s,1H), 7.57 (s, 1H), 7.45 (m, 2H), 6.86 (bs, 1H), 4.31 (AB, 2H), 4.04 (m,1H), 3.24 (m, 2H), 2.40 (m, 1H), 1.94 (m, 1H).

H. 4-Hydroxy-3- 3-(S)-(4,6-dichlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using4,6-dichlorobenzo b!thiophene-2-sulfonic acid1-(2-hydroxy-5-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide as thestarting material. The crude product is purified by RP-HPLC eluting witha gradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ10.80 (bs, 1H), 9.00 (bs, 2H), 8.82 (d, 1H),8.62 (bs, 2H), 8.29 (s, 1H), 7.57 (m, 2H), 7.48 (m, 2H), 6.97 (d, 1H),4.30 (AB, 2H), 4.26 (m, 1H), 3.22 (m, 2H), 2.23 (m, 1H), 1.74 (m, 1H).FAB MS, M+H!⁺ =513, 515, Cl₂ pattern.

EXAMPLE 18 3- 3-(S)-(6-Fluorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 1-Fluoro-3-(2,2-dimethoxy-ethyl-sulfanyl)-benzene

The title compound is prepared as described in EXAMPLE 5, Part Asubstituting 3-fluorothiophenol for 3-chlorothiophenol. The crudeproduct is purified by column chromatography eluting with a gradient ofhexanes to 10% EtOAc/hexanes to afford the title compound as an oil.

¹ H NMR (CDCl₃, 300 MHz) δ7.21 (m, 1H), 7.09 (m, 2H), 6.82 (m, 2H), 4.51(m, 1H), 3.09 (s, 31H), 3.07 (s, 3H).

B. 6-Fluorobenzo b!thiophene

The title compound is prepared as described in EXAMPLE 5, Part Bsubstituting 1-fluoro-3-(2,2-dimethoxy-ethyl-sulfanyl)-benzene for1-chloro-3-(2,2-dimethoxy-ethyl-sulfanyl)-benzene. The crude product ispurified by column chromatography eluting with hexanes to afford thetitle compound as a white solid. EI MS, M!⁺ =152.

C. 6-Fluorobenzo b!thiophene-2-sulfonyl chloride

The title compound is prepared as described in EXAMPLE 1, Part Dsubstituting 6-fluorobenzo b!thiophene for thianaphthalene. The crudeproduct is purified by column chromatography eluting with hexanes toyield the title compound as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.08 (s, 1H), 7.94 (dd, 1H), 7.58 (dd, 1H),7.23 (dt, 1H).

D. 6-Fluorobenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared as described in EXAMPLE 17, Part G,substituting 3- (3-(S)-amino-2-oxo-pyrrolidin-1-yl)-methyl!-benzonitrilefor 3-(3-(S)-amino-2-oxo-pyrrolidin-1-yl)-methyl!-4-hydroxy-benzonitrile andsubstituting 6-fluorobenzo b!thiophene-2-sulfonyl chloride for4,6-dichlorobenzo b!thiophene-2-sulfonyl chloride. The crude product istriturated with Et₂ O to give the product as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ7.85 (s, 1H), 7.78 (m, 1H), 7.64 (m, 2H), 7.49(m, 1H), 7.42 (m, 3H), 7.14 (dt, 1H), 4.42 (AB, 2H), 4.02 (t, 1H), 3.18(m, 2H), 2.60 (m, 1H), 2.12 (m, 1H).

E. 3- 3-(S)-(6Fluorobenzob!thiophene-2sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using6-fluorobenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide as the startingmaterial. The crude product is purified by RP-HPLC eluting with agradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.25 (bs, 4H), 8.63 (d, 1H), 8.03 (s, 1H),8.00 (m, 2H), 7.63 (m, 1H), 7.51 (m, 3H), 7.31 (dt, 1H), 4.38 (AB, 2H),4.28 (m, 1H), 3.11 (m, 2H), 2.13 (m, 1H), 1.68 (m, 1H). FAB MS, M+H!⁺=447. Elemental analysis calculated with 1.0 mole of H₂ O: C=45.67%,H=3.71%, N=9.68%; found: C=45.52%, H=3.95%, N=9.31%.

EXAMPLE 19 4-Amino-3- 3-(S)-(6-fluorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-yl methyl!-benzamidinetrifluoroacetate

A. 6-Fluorobenzo b!thiophene-2-sulfonic acid1-(2-amino-5-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared as described in EXAMPLE 17, Part G,substituting 3-(3-(S)-amino-2-oxo-pyrrolidin-1-yl)-methyl!-4-amino-benzonitrile for 3-(3-(S)-amino-2-oxo-pyrrolidin-1-yl)-methyl!-4-hydroxy-benzonitrile andsubstituting 6-fluorobenzo b!thiophene-2-sulfonyl chloride for4,6-dichlorobenzo b!thiophene-2-sulfonyl chloride. The crude product istriturated with Et₂ O to give the product as a white solid.

FAB MS, M+H!⁺ =445.

B. 4-Amino-3- 3-(S)-(6-fluorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using6-fluorobenzo b!thiophene-2-sulfonic acid1-(2-amino-5-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide as thestarting material. The crude product is purified by RP-HPLC eluting witha gradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 70% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ8.74 (bs, 2H), 8.68 (d, 1H), 8.40 (bs, 2H),8.01 (m, 3H), 7.53 (d, 1 H), 7.36 (m, 2H), 6.72 (d, 1H), 6.20 (bs, 2H),4.28 (m, 1H), 4.26 (AB, 2H), 3.12 (m, 2H), 2.09 (m, 1H), 1.60 (m, 1H).FAB MS, M+H!⁺ =462.

EXAMPLE 20 4-Hydroxy-3- 3-(S)-(6-fluorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 6-Fluorobenzo b!thiophene-2-sulfonic acid1-(2-hydroxy-5-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared as described in EXAMPLE 17, Part G,substituting 6-fluorobenzo b!thiophene-2-sulfonyl chloride for4,6-dichlorobenzo b!thiophene-2-sulfonyl chloride. The crude product istriturated with Et₂ O to give the product as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ7.85 (s, 1H), 7.82 (m, 1H), 7.63 (m, 2H), 7.47(m, 2H), 7.22 (dt, 1H), 6.93 (d, 1H), 4.32 (AB, 2H), 4.20 (t, 1H), 3.32(m, 2H), 2.09 (m, 1H).

B. 4-Hydroxy-3- 3-(S)-(6-fluorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using6-fluorobenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide as the startingmaterial. The crude product is purified by RP-HPLC eluting with agradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 70% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ10.87 (bs, 1H), 9.00 (bs, 2H), 8.68 (bs,2H), 8.60 (d, 1H), 8.01 (s, 1H), 8.00 (m, 2H), 7.54 (d, 1H), 7.38 (m,2H), 6.92 (m, 1H) 4.30 (AB, 2H), 4.21 (m, 1H), 3.15 (m, 2H), 2.18 (m,1H), 1.67 (m, 1H). FAB MS, M+H!⁺ =463.

EXAMPLE 21 4-Amino-3- 3-(S)-(4-chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 4-Chlorobenzo b!thiophene-2-sulfonic acid1-(2-amino-5-Cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared as described in EXAMPLE 17, Part G,substituting 3-(3-(S)-amino-2-oxo-pyrrolidin-1-yl)-methyl!-4-amino-benzonitrile for 3-(3-(S)-amino-2-oxo-pyrrolidin-1-yl)-methyl!-4-hydroxy-benzonitrile andsubstituting 4-chlorobenzo b!thiophene-2-sulfonyl chloride for4,6-dichlorobenzo b!thiophene-2-sulfonyl chloride. The crude product istriturated with Et₂ O to give the product as a white solid.

FAB MS, M+H!⁺ =461, 463, Cl pattern.

B. 4-Amino-3- 3-(S)-(4-chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using4-chlorobenzo b!thiophene-2-sulfonic acid1-(2-amino-5-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide as thestarting material. The crude product is purified by RP-HPLC eluting witha gradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 70% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ8.79 (d, 1H), 8.75 (bs, 2H, 8.36 (bs, 2H),8.07 (d, 1H), 8.03 (s, 1H), 7.52 (d, 1H), 7.50 (m, 2H), 7.37 (m, 2H),6.70 (d, 1H), 6.15 (bs, 2H), 4.27 (m, 1H), 4.13 (AB, 1H), 3.14 (m, 2H),2.13 (m, 1H), 1.60 (m, 1H). FAB MS, M+H!⁺ =478, 480, Cl pattern.

EXAMPLE 22 4Hydroxy-3- 3-(S)-(4-chlorobenzob!thiophene-2-sulfamylamino)2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 4-Chlorobenzo b!thiophene-2-sulfonic acid1-(2-hydroxy-5-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared as described in EXAMPLE 17, Part G,substituting 4-chlorobenzo b!thiophene-2-sulfonyl chloride for4,6-dichlorobenzo b!thiophene-2-sulfonyl chloride. The crude product istriturated with Et₂ O to give the product as a white solid.

FAB MS, M+H!⁺ =462, 464, Cl pattern.

B. 4-Hydroxy-3- 3-(S)-(4-Chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using4-chlorobenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide as the startingmaterial. The crude product is purified by RP-HPLC eluting with agradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 70% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ10.87 (bs, 1H), 9.01 (bs, 2H), 8.74 (d, 2H),8.60 (bs, 1H), 8.08 (d, 1H), 8.02 (s, 1 H), 7.53 (m, 3H), 7.38 (m, 2H),6.92 (d, 1H), 4.30 (AB, 2H), 4.27 (m, 1H), 3.21 (m, 2H), 2.20 (m, 1H),1.72 (m, 1H). FAB MS, M+H!⁺ =479, 481, Cl pattern.

EXAMPLE 23 3- 3-(S)-(4-Chloro-thieno3,2-c!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 3-Thiophene-2-yl-acrylic acid

To a solution of 2-thiophene carboxaldehyde (10 g, 89 mmol), in 300 mLof CH₂ Cl₂ is added methyl(triphenylphosphorylidene)acetate. Thesolution is stirred for 48 h. After this time, the solution isconcentrated. The residue is dissolved in 1000 mL of H₂ O:MeOH:THF(1:1:1). To the resulting solution is added lithium hydroxide (22.9 g,547 mmol). The solution is stirred for 3 h. After this time, thesolution is concentrated to 1/3 its volume. The remaining solution iswashed with EtOAc. The aqueous solution is acidified to pH=5 with 1NHCl. A white precipitate forms. The solid is collected by filtration andis then dried over P₂ O₅ under vacuum.

¹ H NMR (CDCl₃, 300 MHz) δ7.90 (d, 1H), 7.42 (d, 1H), 7.27 9d, 1H), 7.08(d, 1H), 6.22 (d, 1H).

B. 5H-Thieno 3,2-c!pyridin-4-one

To a solution of 3-thiophene-2-yl-acrylic acid (7.39 g, 47.9 mmol) in200 mL of acetone is added triethylamine (4.9 g, 47.9 mmol). Theresulting solution is cooled to 0° C. and ethyl chloroformate (5.7 g,52.8 mmol) is added dropwise. After 2 h, sodium azide (4.67 g, 71.9mmol) in 25 mL of H₂ O is then added. The solution is stirred for 1.5 hat 0° C. After this time, the solution is poured into 300 mL of H₂ O. Awhite precipitate forms which is collected by filtration. The resultingsolid is dried over P₂ O₅ under vacuum. The solid is suspended in 20 mLof diphenyl ether. This solution is added dropwise to a solution oftributylamine (6.9 g, 37.4 mmol) in 200 ml of diphenyl ether at 190° C.After 2 h, the solution is cooled to ambient temperatures. The solutionis diluted with 1000 mL of hexanes and is cooled to 0° C. The resultingsolid is collected by filtration and the solid is washed with hexanes.The title compound (3.6 g, 23.8 mmol) is obtained as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ6.71 (m, 2H), 6.42 (d, 1H), 6.13 (d, 1H).

C. 4-Chloro-thieno 3,2-c!pyridine

5H-Thieno 3,2-c!pyridin-4-one (1.0 g, 6.62 mmol) is dissolved in 30 mLof phosphorous oxy chloride. The solution is heated to 100° C. After 4h, the solution is concentrated. The residue is dissolved in CH₂ Cl₂.The resulting solution is washed with water and saturated NaCl. Theorganic layer is dried over MgSO₄, filtered and concentrated. The crudeproduct is purified by column chromatography eluting with a gradient of40% CH₂ Cl₂ /hexanes to 60% CH₂ Cl₂ /hexanes. The title compound (1.0 g,5.8 mmol) is obtained as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.24 (d, 1H), 7.76 (d, 1H), 7.66 (m, 2H). EIMS, M!⁺ =169, 171, Cl pattern.

D. 4-Chloro-thieno 3,2-c!pyridine-2-sulfonyl chloride

The title compound is prepared as described in EXAMPLE 1, Part Dsubstituting the 4-chloro-thieno 3,2-c!pyridine for thianaphthalene. Thecrude product is purified by column chromatography eluting with agradient of 40% CH₂ Cl₂ /hexanes to 60% CH₂ Cl₂ /hexanes. The titlecompound is obtained as a white solid solid. ¹ H NMR (CDCl₃, 300 MHz)δ8.39 (d, 1H), 8.28 (s, 1H), 7.72 (d, 1H).

E. 4-Chloro-thieno 3,2-c!pyridine-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-yl!-amide

The title compound is prepared as described in EXAMPLE 17, Part G,substituting 3-(3-(S)-amino-2oxo-pyrrolidin-1-yl)-methyl-benzonitrilefor 3-(3-(S)-amino-2-oxo-pyrrolidin-1-yl)-methyl!-4-hydroxy-benzonitrile andsubstituting 4-chloro-thieno 3,2-c!pyridine-2-sulfonyl chloride for4,6-dichlorobenzo b!thiophene-2-sulfonyl chloride. The crude product ispurified by column chromatography eluting with a gradient of 10% CH₂ Cl₂/EtOAc to 20% CH₂ Cl₂ /EtOAc. The title compound is obtained as a whitesolid solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.34 (m, 1H), 8.11 (s, 1H), 7.73 (m, 1H), 7.58(m, 1H), 7.40 (m, 3H), 5.62 (bs, 1H), 4.46 (AB, 2H), 3.97 (m, 1H), 3.27(m, 2H), 2.68 (m, 1H), 2.12 (m, 1H).

F. 3- 3-(S)-(4-Chloro-thieno3,2-c!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using4-chlorobenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide as the startingmaterial. The crude product is purified by RP-HPLC eluting with agradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 70% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.28 (bs, 2H), 9.18 (bs, 2H), 9.06 (d, 1H),8.38 (d, 1H), 8.22 (d, 1H), 8.08 (s, 1H), 7.68 (m, 1H), 7.52 (m, 3H),4.41 (AB, 2H), 4.33 (m, 1H), 3.14 (m, 2H), 2.21 (m, 1H), 1.72 (m, 1H).FAB MS, M+H!⁺ =464, 466, Cl pattern.

EXAMPLE 24 4Hydroxy-3- 3-(S)-(4-chloro-thieno3,2-c!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 4-Chloro-thieno 3.2-c!pyridine-2-sulfonic acid1-(2-hydroxy-5-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared as described in EXAMPLE 17, Part G,substituting 4-chloro-thieno 3,2-c!pyridine-2-sulfonyl chloride for4,6-dichlorobenzo b!thiophene-2-sulfonyl chloride. The crude product ispurified by column chromatography eluting with a gradient of 1% MeOH/CH₂Cl₂ to 4% MeOH/CH₂ Cl₂. The title compound is obtained as a white solidsolid.

¹ H NMR (CDCl₃, 300 MHz) δ9.13 (s, 1H), 8.38 (d, 1H), 8.09 (s, 1H), 7.72(d, 1H), 7.52 (dd, 1H), 7.38 (s, 1H), 6.96 (d, 1H), 5.44 (d, 1H), 4.32(AB, 2H), 4.10 (m, 1H), 3.48 (m, 2H), 2.74 (m, 1H), 2.18 (m, 1H).

B. 4-Hydroxy-3- 3-(S)-(4-chloro-thieno3,2-c!pyridine-2-sulfonylamino-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using4-chlorobenzo b!thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide as the startingmaterial. The crude product is purified by RP-HPLC eluting with agradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 70% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ10.91 (bs, 1H), 9.04 (bs, 2H), 8.93 (d, 1H),8.78 (bs, 2H), 8.40 (d, 1H), 8.21 (d, 1H), 8.08 (s, 1H), 7.62 (m, 1H),7.41 (s, 1H), 6.97 (d, 1H), 4.31 (AB, 2H), 4.30 (m, 1H), 3.22 (m, 2H),2.26 (m, 1H), 1.78 (m, 1H). FAB MS, M+H!⁺ =480, 482, Cl pattern.

EXAMPLE 25 3- 3-(S)-(5-Chlorothieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 5-Chlorothieno 3,2-b!pyridine

The title compound is prepared from 3-acetyl thiophene according to theprocedure described in J. Chem. Soc., Perkin Trans. I, 1981, 1531. Thecrude product is purified by column chromatography eluting with agradient of 10% EtOAc/hexanes to 20% EtOAc/hexanes to afford a whitesolid.

¹ H NMR (CDCl₃, 300 MHz) δ8.12 (d, 1H), 7.80 (d, 1H), 7.51 (d, 1H), 7.28(d, 1H), EI MS, M!⁺ =169, 171, Cl pattern.

B. 5-Chlorothieno 3,2-b!pyridine-2-sulfonyl chloride

The title compound is prepared as described in EXAMPLE 1, Part D using5-chlorothieno 3,2-b!pyridine in place of thianaphthalene. The crudeproduct is obtained as a white solid and is of sufficient purity to beused in the subsequent step.

¹ H NMR (CDCl₃, 300 MHz) δ8.25 (s, 1H), 8.23 (d, 1H), 7.53 (d, 1H). EIMS, M!⁺ =267, 269, Cl pattern.

C. 5-Chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared from3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)-benzonitrile hydrochlorideas described in EXAMPLE 1, Part E using 5-chlorothieno3,2-b!pyridine-2-sulfonyl chloride in place of benzob!thiophene-2sulfonyl chloride. The product is obtained as a whitesolid.

¹ H NMR (CDCl₃, 300 MHz) δ8.18 (d, 1H), 8.03 (s, 1H), 7.59 (m, 1H), 7.47(d, 1H), 7.45 (m, 2H), 7.42 (d, 1H), 5.95 (bs, 1H), 4.48 (s, 2H), 3.99(m, 1H), 3.28 (m, 2H), 2.67 (m, 1H),2.14 (m, 1H).

D. 3- 3-(S)-(5-Chlorothieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

5-Chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide is converted to thetitle compound as described in EXAMPLE 1, Part F. The crude product ispurified by RP-HPLC eluting in a gradient of 10% CH₃ CN/H₂ O (0.1% TFA)to 70% CH₃ CN/H₂ O (0.1% TFA) and the appropriate product fractions arelyophilized to provide the title compound as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.30 (bs, 2H), 9.17 (bs, 2H), 8.93 (d, 1H),8.67 (d, 1H), 8.11 (s, 1H), 7.69 (m, 1H), 7.64 (d, 1H), 7.55 (m, 3H),4.43 (AB, 2H), 4.33 (m, 1H), 3.16 (m, 2H), 2.22 (m, 1H), 1.73 (m, 1H).FAB MS, M+H!⁺ =464, 466, Cl pattern. Elemental analysis calculated with1.0 mol H₂ O; C=42.35%, H=3.55%, N=11.76%, found: C=42.34%, H=3.30%,N=11.39%.

EXAMPLE 26 3- 3-(S)-(Thieno3,2-b!pyridine-2-sulfonylamino-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinebistrifluoroacetate

To a solution of 3- 3-(S)-(5-chlorothieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate (0.21 g, 0.36 mmol) in 10 mL of 1:1 benzene/MeOH isadded excess solid KOH (0.25 g) and a catalytic amount of 10% palladiumon activated carbon. The heterogenous mixture is hydrogenated at roomtemperature on a Parr apparatus under 70 p.s.i. of H₂ for 5 days. Thereaction mixture is filtered through a pad of Celite, washed with MeOH,and the filtrate is concentrated in vacuo. The crude product is purifiedby RP-HPLC eluting in a gradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 60%CH₃ CN/H₂ O (0.1% TFA) and the appropriate product fractions arelyophilized to provide the title compound (0.14 g, 0.21 mmol) as a whitesolid.

¹ H NMR (DMSO-d₆, 500 MHz) δ9.56 (bs, 2H), 9.35 (bs, 2H), 8.84 (d, 1H),8.80 (d, 1H), 8.61 (d, 1H), 8.14 (s, 1H), 7.72 (dt, 1H), 7.61 (s, 1H),7.57 (dd, 1H), 7.56 (s, 1H), 7.54 (dd, 1), 4.44 (AB, 2H), 4.34 (m, 1H),3.17 (m, 2H), 2.21 (m, 1H), 1.74 (m, 1H). IS MS, M+H!⁺ =430.

EXAMPLE 27 3- 3-(S)-(5-Chlorothieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-hydroxybenzamidinetrifluoroacetate

5-Chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide (0.20 g, 0.45 mmol)is dissolved in 10 mL of MeOH. The solution is cooled to 0° C. and HClgas is bubbled through the solution for 5 min. The reaction mixture isallowed to warm to room temperature and is stirred for 16 h. At thistime, the solution is concentrated in vacuo and pumped dry under highvacuum. To a solution of the residue dissolved in 20 mL of MeOH is addedhydroxylamine hydrochloride (0.78 g, 11.2 mmol) followed bytriethylamine (2.23 g, 22.0 mmol). The resulting solution is heated atreflux for 2 h. The solution is concentrated and the residue is purifiedby RP-HPLC eluting with a gradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 70%CH₃ CN/H₂ O (0.1% TFA) and the appropriate fractions are lyophilized togive the title compound (0.17 g, 0.29 mmol) as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ8.94 (d, 1H), 8.78 (bs, 1H), 8.67 (d, 1H),8.10 (s, 1H), 7.64 (d, 1H), 7.57 (m, 2H), 7.50 (m, 2H), 4.43 (AB, 2H),4.33 (m, 1H), 3.16 (m, 2H), 2.22 (m, 1H), 1.71 (m, 1H). FAB MS, M+H!⁺=480, 482, Cl pattern. Elemental analysis calculated with 1.4 mol H₂ Ocal. C=40.74%, H=3.55%, N=11.31%, found: C=40.75%, H=3.20%, N=10.86%.

EXAMPLE 28 3-{3-(S)- (5-Chlorothieno3,2-b!pyridine-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl}benzamidine trifluoroacetate

A. 5-Chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-methylamide

5-Chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide (0.25 g, 0.56 mmol)is dissolved in 6 mL of DMF and cooled to 0° C. To the solution is addedmethyl iodide (0.40 g, 2.82 mmol) and sodium hydride (25 mg of a 60%dispersion in mineral oil, 0.62 mmol). The reaction mixture is allowedto warm to room temperature and is stirred for 2 h. At this time, thesolution is diluted with and EtOAc and the layers are separated. Theorganic layer is washed with 1N HCl, H₂ O, saturated NaHCO₃ solution andsaturated NaCl. The organic phase is then dried over MgSO₄, filtered andconcentrated to give the title compound (0.25 g, 0.54 mmol) as a solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.15 (d, 1H), 8.08 (s, 1H), 7,60 (m, 1H), 7.45(m, 3H), 7.40 (d, 1H), 4.92 (m, 1H), 4.42 (AB, 2H), 3.24 (m, 2H), 2.92(s, 3H), 2.43 (m, 1H), 2.06 (m, 1H).

B. 3-{3-(S)- (5-Chlorothieno3,2-b!pyridine-2sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl}-benzamidinetrifluoroacetate

5-Chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-methylamide is converted tothe title compound as described in EXAMPLE 1, Part F. The crude productis purified by RP-HPLC eluting in a gradient of 10% CH₃ CN/H₂ O (0.1%TFA) to 60% CH₃ CN/H₂ O (0.1% TFA) and the appropriate product fractionsare lyophilized to afford the title compound as a white solid.

¹ H NMR (DMSO-d₆, 500 MHz) δ9.32 (bs, 2H), 9.14 (bs, 2H), 8.69 (d, 1H),8.24 (s, 1H), 7.70 (m, 1H), 7.66 (d, 1H), 7.58 (m, 2H), 7.56 (s, 1 H),5.00 (m, 1 H), 4.44 (AB, 2H), 3.24 (m, 1H), 3.19 (m, 1H), 2.83 (s, 3H),2.17 (m, 1H), 1.96 (m, 1H). FAB MS, M+H!⁺ =478, 480, Cl pattern.Elemental analysis calculated with 1.3 mol H₂ O: C=42.91%, H=3.87%,N=11.37%, found: C=42.90%, H=3.52%, N=11.07%.

EXAMPLE 29 3- 3-(S)-(6-Chlorothieno2,3-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 2-Bromo-6-chlorothieno 2,3-b!pyridine

The title compound is prepared from 2-bromo-5-acetyl thiophene accordingto the procedure described in J. Chem. Soc., Perkin Trans. I, 1981,1531. The crude product is purified by column chromatography elutingwith 2% EtOAc/hexanes to afford a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ7.89 (d, 1H), 7.28 (d, 1H), 7.27 (d, 1H).

B. 6-Chlorothieno 2,3-b!pyridine-2-sulfonyl chloride

The title compound is prepared as described in EXAMPLE 1, Part D using2-bromo-6-chlorothieno 2,3-b!pyridine in place of thianaphthalene. Thecrude product is obtained as a white solid and is of sufficient purityto be used in the subsequent step.

¹ H NMR (CDCl₃, 300 MHz) δ8.22 (d, 1H), 8.09 (s, 1H), 7.52 (d, 1H). EIMS, M!⁺ =267, 269, Cl pattern.

C. 6-Chlorothieno 2,3-b!pyridine-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared from3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)-benzonitrile hydrochlorideas described in EXAMPLE 1, Part E using 6-chlorothieno2,3-b!pyridine-2-sulfonyl chloride in place of benzob!thiophene-2-sulfonyl chloride. The crude product is purified by columnchromatography eluting with a gradient of 10% EtOAc/CH₂ Cl₂ to 25%EtOAc/CH₂ Cl₂ to afford a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.12 (d, 1H), 7.89 (s, 1H), 7.57 (m, 1H), 7.50(s, 1H), 7.45 (m, 2H), 7.42 (d, 1H), 6.03 (bs, 1 H), 4.48 (AB, 2H), 4.05(m, 1H), 3.26 (m, 2H), 2.68 (m, 1H), 2.15 (m, 1H).

D. 3- 3-(S)-(6-Chlorothieno2,3-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

6-Chlorothieno 2,3-b!pyridine-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide is converted to thetitle compound as described in EXAMPLE 1, Part F. The crude product ispurified by RP-HPLC eluting in a gradient of 10% CH₃ CN/H₂ O (0.1% TFA)to 60% CH₃ CN/H₂ O (0.1% TFA) and the appropriate product fractions arelyophilized to afford the title compound as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.29 (bs, 2H), 9.15 (bs, 2H), 8.88 (d, 1H),8.48 (d, 1H), 8.08 (s, 1H), 7.68 (m, 2H), 7.56 (m, 3H), 4.42 (AB, 2H),4.30 (m, 1H), 3.16 (m, 2H), 2.22 (m, 1H), 1.73 (m, 1 H). FAB MS, M+H!⁺=464, 466, Cl pattern. Elemental analysis calculated with 1.4 mol H₂ Ocal. C=41.78%, H=3.65%, N=11.60%, found: C=41.78%, H=3.28%, N=11.16%.

EXAMPLE 30 3- 3-(S)-(Thieno2,3-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinebistrifluoroacetate

3- 3-(S)-(6-Chlorothieno 2,3-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidine trifluoroacetate is convertedto the title compound as described in EXAMPLE 26. The reaction mixtureis hydrogenated at room temperature for 3 days. The crude product ispurified by RP-HPLC eluting in a gradient of 10% CH₃ CN/H₂ O (0.1% TFA)to 80% CH₃ CN/H₂ O (0.1% TFA) and the appropriate product fractions arelyophilized to provide the title compound as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.22 (bs, 2H), 9.04 (bs, 2H), 8.73 (d, 1H),8.64 (m, 1H), 8.37 (d, 1H), 7.99 (s, 1H), 7.62 (m, 1H), 7.50 (m, 4H),4.36 (AB, 2H), 4.21 (m, 1H), 3.08 (m, 2H), 2.13 (m, 1H), 1.67 (m, 1H).IS MS, M+H!⁺ =430.

EXAMPLE 31 3-{3-(S)- (6-Chlorothieno2,3-b!pyridine-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl}-benzamidinetrifluoroacetate

A. 6-Chlorothieno 2,3-b!pyridine-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S) -yl!-methylamide

The title compound is prepared as described in EXAMPLE 28, Part A using6-chlorothieno 2,3-b!pyridine-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide in place of5-chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide. The crude product ispurified by column chromatography eluting with a gradient of 5%EtOAc/CH₂ Cl₂ to 10% EtOAc/CH₂ Cl₂ to yield a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.13 (d, 1H), 7.97 (s, 1H), 7.60 (m, 1H), 7.48(m, 3H), 7.41 (d, 1H), 4.94 (m, 1H), 4.44 (AB, 2H), 3.25 (m, 2H), 2.93(s, 3H), 2.44 (m, 1H), 2.09 (m, 1H).

B. 3-{3-(S)- (6-Chlorothieno2,3-b!pyridine-2sulfonyl-methylamino!-2-oxo-pyrrolidin-1-ylmethyl}-benzamidinetrifluoroacetate

6-Chlorothieno 2,3-b!pyridine-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-methylamide is converted tothe title compound as described in EXAMPLE 1, Part F. The crude productis purified by RP-HPLC eluting in a gradient of 10% CH₃ CN/H₂ O (0.1%TFA) to 80% CH₃ CN/H₂ O (0.1% TFA) and the appropriate product fractionsare lyophilized to afford the title compound as a white solid.

¹ H NMR (DMSO-d₆, 500 MHz) δ9.30 (bs, 2H), 9.03 (bs, 2H), 8.49 (d, 1H),8.17 (s, 1H 7.70 (d, 1H), 7.68 (m, 1H), 7.55 (m, 3H), 4.96 (m, 1H), 4.43(AB, 2H), 3.19 (m, 2H), 2.81 (s, 3H), 2.15 (m, 1H), 1.97 (m, 1H). FABMS, M+H!⁺ =478, 480, Cl pattern.

EXAMPLE 32 4-Hydroxy-3- 2-oxo-3-(S)-(5-chlorothieno3,2-b!pyridine-2-sulfonylamino)-pyrrolidin-1-ylmethyl}-benzamidinetrifluoroacetate

A. 5-Chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(5-cyano-2-hydroxybenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared from4-hydroxy-3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)-benzonitrilehydrochloride as described in EXAMPLE 17, Part F using 5-chlorothieno3,2-b!pyridine-2-sulfonyl chloride in place of 4,6-dichlorobenzob!thiophene-2-sulfonyl chloride. The product is obtained as a whitesolid.

¹ H NMR (CDCl₃, 300 MHz) δ9.40 (bs, 1H), 8.16 (d, 1H), 8.02 (s, 1H),7.52 (dd, 1H), 7.43 (d, 1H), 7.38 (s, 1H), 6.96 (d, 1H), 5.65 (bs, 1H),4.31 (AB, 2H), 4.10 (m, 1H), 3.46 (m, 2H), 2.68 (m, 1H), 2.15 (m, 1H).

B. 4Hydroxy-3- 2-oxo-3-(S)-(5chlorothieno3,2-b!pyridine-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

5-Chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(5-cyano-2-hydroxybenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide isconverted to the title compound as described in EXAMPLE 1, Part F. Thecrude product is purified by RP-HPLC eluting in a gradient of 10% CH₃CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA) and the appropriateproduct fractions are lyophilized to afford the title compound as awhite solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.03 (bs, 2H), 8.93 (d, 1H), 8.75 (bs, 2H),8.67 (d, 1H), 8.11 (s, 1H), 7.65 (d, 1H), 7.59 (dd, 1H), 7.41 (s, 1H),6.97 (d, 1H), 4.33 (AB, 2H), 4.29 (m, 1H), 3.21 (m, 2H), 2.22 (m, 1H),1.74 (m, 1H). IS MS, M+H!⁺ =480, 482, Cl pattern. Elemental analysiscalculated with 1.2 mol H₂ O: C=41.03%, H=3.49%, N=11.39%, found:C=41.02%, H=3.24%, N=10.86%.

EXAMPLE 33 4-Hydroxy-3- 2-oxo-3-(S)-(thieno3,2-b!pyridine-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinebistrifluoroacetate

5- 3-(S)-(5-Chlorothieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-2-hydroxy-benzamidinetrifluoroacetate is converted to the title compound as described inEXAMPLE 26. The reaction mixture is hydrogenated at room temperature for7 days. The crude product is purified by RP-HPLC eluting in a gradientof 10% CH₃ CN/H₂ O (0.1% TFA) to 60% CH₃ CN/H₂ O (0.1% TFA) and theappropriate product fractions are lyophilized to provide the titlecompound as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.03 (bs, 2H), 8.85 (bs, 2H), 8.83 (d, 1H),8.78 (d, 1H), 8.60 (d, 1H), 8.12 (s, 1H), 7.61 (d, 1H), 7.53 (dd, 1H),7.42 (s, 1H), 6.97 (d, 1H), 4.33 (AB, 2H), 4.28 (m, 1H), 3.22 (m, 2H),2.22 (m, 1H), 1.76 (m, 1H). FAB MS, M+H!⁺ =446.

EXAMPLE 34 4Hydroxy-3- 2-oxo-3-(S)-(5-chlorothieno3,2-b!pyridine-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-hydroxybenzamidinetrifluoroacetate

5-Chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(5-cyano-2-hydroxybenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide isconverted to the title compound as described in EXAMPLE 27. The crudeproduct is purified by RP-HPLC eluting in a gradient of 10% CH₃ CN/H₂ O(0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA) and the appropriate productfractions are lyophilized to provide the title compound as a whitesolid.

¹ H NMR (DMSO-d₆, 300 MHz) δ8.93 (d, 1H), 8.89 (bs, 1H), 8.68 (d, 1H),8.11 (s, 1H), 7.65 (d, 1H), 7.48 (d, 1H), 7.30 (s, 1H), 6.97 (d, 1H),4.32 (AB, 2H), 4.28 (m, 1H), 3.20 (m, 2H), 2.22 (m, 1H), 1.73 (m, 1H).FAB MS, M+H!⁺ =496, 498, Cl pattern.

EXAMPLE 35 4Amino-3- 2-oxo-3-(S)-(5-chlorothieno3,2-b!pyridine2sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 5-Chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(2-amino-5-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared from4-amino-3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)-benzonitriledihydrochloride as described in EXAMPLE 17, Part F using 5-chlorothieno3,2-b!pyridine-2-sulfonyl chloride in place of 4,6-dichlorobenzob!thiophene-2-sulfonyl chloride. The crude product is purified by columnchromatography eluting with a gradient of 10% EtOAc/CH₂ Cl₂ to 25%EtOAc/CH₂ Cl₂ to afford a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.16 (d, 1H), 8.03 (s, 1H), 7.44 (d, 1H), 7.38(dd, 1H), 7.28 (dd, 1 H), 6.60 (d, 1H), 5.55 (bs, 1H), 4.86 (bs, 2H),4.29 (AB, 2H), 4.00 (m, 1H), 3.26 (m, 2H), 2.66 (m, 1H), 2.09 (m, 1H).

B. 4-Amino-3- 2-oxo-3-(S)-(5-chlorothieno3,2-b!pyridine-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

5-Chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(2-amino-5-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide is convertedto the title compound as described in EXAMPLE 1, Part F. The crudeproduct is purified by RP-HPLC eluting in a gradient of 10% CH₃ CN/H₂ O(0.1% TFA) to 70% CH₃ CN/H₂ O (0.1% TFA) and the appropriate productfractions are lyophilized to afford the title compound as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ8.94 (d, 1H), 8.80 (bs, 2H), 8.68 (d, 1H),8.40 (bs, 2H), 8.11 (s, 1H), 7.65 (d, 1H), 7.54 (dd, 1H), 7.44 (s, 1H),6.72 (d, 1H), 6.19 (bs, 2H), 4.33 (m, 1H), 4.20 (AB, 2H), 3.20 (m, 2H),2.22 (m, 1H), 1.70 (m, 1H). FAB MS, M+H!⁺ =479, 481, Cl pattern.Elemental analysis calculated with 2.0 mol H₂ O cal. C=40.00%, H=3.86%,N=13.34%, found: C=40.03%, H=3.19%, N=12.67%.

EXAMPLE 36 4-Amino-3- 2-oxo-3-(S)-(thieno3,2-b!pyridine-2-sulfonylamino-pyrrolidin-1-ylmethyl!-benzamidinebistrifluoroacetate

4-Amino-3- 3-(S)-(5-chlorothieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate is converted to the title compound as described inEXAMPLE 26. The reaction mixture is hydrogenated at room temperature for7 days. The crude product is purified by RP-HPLC eluting in a gradientof 10% CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA) and theappropriate product fractions are lyophilized to provide the titlecompound as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ8.85 (d, 1H), 8.78 (bs, 3H), 8.60 (d, 1H),8.39 (bs, 2H), 8.11 (s, 1H), 7.53 (m, 1H), 7.52 (d, 1H), 7.42 (s, 1H),6.71 (d, 1H), 6.17 (bs, 1H), 4.31 (m, 1H), 4.19 (AB, 1H), 3.19 (m, 2H),2.20 (m, 1H), 1.69 (m, 1H). FAB MS, M+H!⁺ =445.

EXAMPLE 37 4- 3-(S)-(5-Chlorothieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-thiophene-2-carboxamidinetrifluoroacetate

A. 5-Chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(5-cyanothiophen-3-ylmethyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared from4-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)-thiophene-2carbonitrilehydrochloride as described in EXAMPLE 1, Part E using 5-chlorothieno3,2-b!pyridine-2-sulfonyl chloride in place of benzob!thiophene-2-sulfonyl chloride. The product is triturated with Et₂O/CH₂ Cl₂ /hexanes to give a white solid.

¹ H NMR (CDCl₃ +CD₃ OD, 300 MHz) δ8.20 (d, 1H), 8.03 (s, 1H), 7.50 (s,1H), 7.48 (s, 1H), 7.43 (d, 1H), 4.41 (AB, 2H), 4.08 (m, 1 H), 3.27 (m,2H), 2.58 (m, 1H), 2.05 (m, 1H).

B. 4- 3-(S)-(5-Chlorothieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-thiophene-2-carboxamidinetrifluoroacetate

5-Chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(5-cyanothiophen-3-ylmethyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide isconverted to the title compound as described in EXAMPLE 1, Part F. Thecrude product is purified by RP-HPLC eluting in a gradient of 10% CH₃CN/H₂ O (0.1% TFA) to 60% CH₃ CN/H₂ O (0.1% TFA) and the appropriateproduct fractions are lyophilized to afford the title compound as awhite solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.26 (bs, 2H), 9.12 (bs, 2H), 8.92 (d, 1H),8.67 (d, 1H), 8.10 (s, 1H), 7.89 (s, 1H), 7.81 (s, 1H), 7.63 (d, 1H),4.37 (AB, 2H), 4.28 (m, 1H), 3.19 (m, 2H), 2.22 (m, 1H), 1.73 (m, 1H).FAB MS, M+H!⁺ =470, 472, Cl pattern. Elemental analysis calculated with2.1 mol H₂ O cal. C=36.70%, H=3.44%, N=11.26%, found: C=36.70%, H=2.78%,N=10.38%.

EXAMPLE 38 4- 3-(S)-(5-Chlorothieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-thiophene-2-hydroxycarboxamidinetrifluoroacetate

5-Chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(5-cyanothiophen-3-ylmethyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide isconverted to the title compound as described in EXAMPLE 27. The crudeproduct is purified by RP-HPLC eluting in a gradient of 10% CH₃ CN/H₂ O(0.1% TFA) to 60% CH₃ CN/H₂ O (0.1% TFA) and the appropriate productfractions are lyophilized to provide the title compound as a whitesolid.

¹ H NMR (DMSO-d₆, 300 MHz) δ8.91 (d, 1H), 8.68 (d, 1H), 8.11 (s, 1H),7.65 (dd, 1H), 7.55 (s, 1H), 7.47 (s, 1H), 4.33 (AB, 2H), 4.28 (m, 1H),3.18 (m, 2H), 2.22 (m, 1H), 1.72 (m, 1H). FAB MS, M+H!⁺ =486, 488, Clpattern. Elemental analysis calculated with 1.5 mol H₂ O cal. C=36.42%,H=3.21%, N=11.18%, found: C=36.43%, H=2.77%, N=10.83%.

EXAMPLE 39 4-{3-(S)-(5-Chlorothieno3,2-b!pyridine-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl}-thiophene-2-carboxamidinetrifluoroacetate

A. 5-Chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(5-Cyanothiophen-3-ylmethyl)-2-oxo-pyrrolidin-3-(S)-yl!-methylamide

The title compound is prepared as described in EXAMPLE 28, Part A using5-chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(5-cyanothiophen-3-ylmethyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide in placeof 5-chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide. The crude product isobtained as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.20 (d, 1H), 8.05 (s, 1H), 7.48 (s, 1H), 7.45(s, 1H), 7.40 (d, 1H), 4.91 (m, 1H), 4.40 (AB, 2H), 3.30 (m, 2H), 2.90(s, 3H), 2.40 (m, 1H), 2.05 (m, 1H).

B. 4-{3-(S)- (5-Chlorothieno3,2-b!pyridine-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl}-thiophene-2-carboxamidinetrifluoroacetate

5-Chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(5-cyanothiophen-3-ylmethyl)-2-oxo-pyrrolidin-3-(S)-yl!-methylamide isconverted to the title compound as described in EXAMPLE 1, Part F. Thecrude product is purified by RP-HPLC eluting in a gradient of 10% CH₃CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA) and the appropriateproduct fractions are lyophilized to provide the title compound as awhite solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.25 (bs, 2H), 8.98 (bs, 2H), 8.70 (d, 1H),8.23 (s, 1H), 7.91 (s, 1H), 7.80 (s, 1H), 7.66 (d, 1H), 4.95 (m, 1H),4.36 (AB, 2H), 3.22 (m, 2H), 2.79 (s, 3H), 2.13 (m, 1H), 1.94 (m, 1H).FAB MS, M+H!+=484, 486, Cl pattern. Elemental analysis calculated with1.3 mol H₂ O: C=38.68%, H=3.50%, N=11.28%, found: C=38.69%, H=2.99%,N=10.72%.

EXAMPLE 40 3-{3-(S)-5-(2-Methylsulfanyl-pyrimidin-4yl)-thiophene-2-sulfonylamino!-2-oxo-pyrrolidin-1-ylmethyl}-benzamidinetrifluoroacetate

A. 3-(3-(S)-Amino-2-oxo-pyrrolidin-1-ylmethyl)-benzamidineditrifluoroacetate

1-(3-Cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-carbamic acid tert-butylester is converted to the title compound as described in EXAMPLE 1, PartF. The crude product is purified by RP-HPLC eluting in a gradient of 2%CH₃ CN/H₂ O (0.1% TFA) to 30% CH₃ CN/H₂ O (0.1% TFA) and the appropriateproduct fractions are lyophilized to afford the title compound as awhite solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.36 (bs, 2H), 9.28 (bs, 2H), 8.47 (bs, 3H),7.75 (m, 1H), 7.65 (s, 1H), 7.62 (m, 2H), 4.55 (AB, 2H), 4.13 (m, 1H),3.33 (m, 2H), 2.40 (m, 1H), 1.94 (m, 1H). FAB MS, M+H!⁺ =233. Elementalanalysis calculated with 0.7 mol H₂ O: C=40.59%, H=4.14%, N=11.83%,found: C=40.58%, H=3.92%, N=11.81%.

B. 3-{3-(S)-5-(2-Methylsulfanyl-pyrimidin-4-yl)-thiophene-2-sulfonylamino!-2-oxo-pyrrolidin-1-ylmethyl}-benzamidinetrifluoroacetate

The title compound is prepared from3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)-benzamidineditrifluoroacetate as described in EXAMPLE 1, Part E using 5-2-(methylthio)-pyrimidin-4-yl!thiophene2-sulfonyl chloride in place ofbenzo b!thiophene-2-sulfonyl chloride. The crude product is purified byRP-HPLC eluting in a gradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃CN/H₂ O (0.1% TFA) and the appropriate product fractions are lyophilizedto provide the title compound as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.28 (bs, 2H), 8.98 (bs, 2H), 8.71 (d, 1H),8.62 (d, 1H), 8.09 (d, 1H), 7.80 (d, 1H), 7.74 (d, 1H), 7.67 (m, 1H),7.57 (m, 3H), 4.43 (AB, 2H), 4.25 (m, 1H), 3.14 (m, 2H), 2.55 (s, 3H),2.18 (m, 1H), 1.70 (m, 1H). FAB MS, M+H!⁺ =503.

EXAMPLE 41 3-{3-(S)-5-(2Methoxy-pyrimidin-4-yl)-thiophene-2-sulfonylamino!-2-oxo-pyrrolidin-1-ylmethyl}-benzamidinetrifluoroacetate

A. 5-(2-Methylsulfanyl-pyrimidin-4-yl)-thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared from3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)-benzonitrile hydrochlorideas described in EXAMPLE 1, Part E using 5-2-(methylthio)-pyrimidin-4-yl!thiophene-2-sulfonyl chloride in place ofbenzo b!thiophene-2-sulfonyl chloride. The crude product is obtained asa white solid and is of sufficient purity to be used in the subsequentstep.

¹ H NMR (CDCl₃, 300 MHz) δ8.55 (d, 1H), 7.70 (d, 1H), 7.65 (d, 1H), 7.60(m, 1H), 7.50 (s, 1H), 7.46 (m, 2H), 7.23 (d, 1H), 5.84 (s, 1H), 4.48(s, 2H), 3.97 (m, 1H), 3.28 (m, 2H), 2.66 (m, 1H), 2.60 (s, 3H), 2.15(m, 1H).

B. 5-(2-Methylsulfinyl-pyrimidin-4-yl)-thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide and5-(2-Methylsulfonyl-pyrimidin-4-yl)-thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

To a solution of5-(2-methylsulfanyl-pyrimidin-4-yl)-thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide (1.48 g, 3.05 mmol)in 30 mL of CHCl₃ at 0° C. is added dropwise a solution of3-chloroperoxybenzoic acid (75%, 1.05 g, 4.58 mmol) in 50 mL of CHCl₃.The resulting solution is allowed to warm to room temperature and isstirred for 16 h. The reaction mixture is diluted with CH₂ Cl₂ andwashed successively with a saturated solution of Na₂ SO₃, 10% Na₂ CO₃and H₂ O. The organic phase is then dried over MgSO₄, filtered andconcentrated in vacuo to give a mixture of the title compounds (1.46 g,2.82 mmol) as a solid which is of sufficient purity to be used in thesubsequent step.

FAB MS, M+H!⁺ =502, 518.

C. 5-(2-Methoxy-pyrimidin-4-yl-thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

A mixture of 5-(2-methylsulfinyl-pyrimidin-4-yl)-thiophene-2-sulfonicacid 1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide and5-(2-methylsulfonyl-pyrimidin-4-yl)-thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide (0.19 g, 0.37 mmol)is dissolved in 10 mL of MeOH and 1 mL of CH₂ Cl₂ and NH₃ gas is bubbledthrough the solution for 5 min. The resulting mixture is heated atreflux for 4 h. After this time, the solution is concentrated in vacuoto give the title compound (0.17 g, 0.36 mmol) as a solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ8.70 (d, 1H), 8.09 (d, 1H), 7.75 (m, 3H),7.66 (s, 1H), 7.55 (m, 3H), 4.42 (AB, 2H), 4.27 (m, 1H), 3.95 (s, 3H),3.15 (m, 2H), 2.17 (m, 1H), 1.68 (m, 1H).

D. 3-{3-(S)-5-(2-Methoxy-pyrimidin-4-yl)-thiophene-2sulfonylamino!-2-oxo-pyrrolidin-1-ylmethyl}-benzamidinetrifluoroacetate

5-(2-Methoxy-pyrimidin-4-yl)-thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide is converted to thetitle compound as described in EXAMPLE 1, Part F. The crude product ispurified by RP-HPLC eluting in a gradient of 10% CH₃ CN/H₂ O (0.1% TFA)to 60% CH₃ CN/H₂ O (0.1% TFA) and the appropriate product fractions arelyophilized to afford the title compound as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.29 (bs, 2H), 9.11 (bs, 2H), 8.70 (d, 1H),8.62 (d, 1H), 8.09 (d, 1H), 7.75 (m, 2H), 7.68 (m, 1H), 7.57 (m, 3H),4.45 (AB, 2H), 4.25 (m, 1H), 3.95 (s, 3H), 3.16 (m, 2H), 2.18 (m, 1H),1.70 (m, 1H). FAB MS, M+H!⁺ =487.

Elemental analysis calculated with 2.5 mol H₂ O: C=42.70%, H=4.39%,N=12.99%, found: C=42.69%, H=3.64%, N=12.28%.

EXAMPLE 42 3-{3-(S)-5(2Amino-pyrimidin-4yl)-thiophene-2-sulfonylamino!-2-oxo-pyrrolidin-1-ylmethyl}-benzamidineditrifluoroacetate

A mixture of 5-(2-methylsulfinyl-pyrimidin-4-yl)-thiophene-2-sulfonicacid 1-(3-cyanobenzyl)-2-oxo-pyrrolidin3-(S)-yl!-amide and5-(2-methylsulfonyl-pyrimidin-4-yl)-thiophene-2sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide (0.20 g, 0.39 mmol)is dissolved in 10 mL of EtOH and NH₃ gas is bubbled through thesolution for 5 min. The resulting mixture is heated at 90° C. for 3 h ina stainless steel Parr high pressure reaction vessel. After this time,the solution is allowed to cool to room temperature and is concentratedin vacuo to give 5-(2-amino-pyrimidin-4-yl)-thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide (0.20 g) as a crudeproduct. FAB MS, M+H!⁺ =455. The crude5-(2-amino-pyrimidin-4-yl)-thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide is converted to thetitle compound as described in EXAMPLE 1, Part F. The crude product ispurified by RP-HPLC eluting in a gradient of 10% CH₃ CN/H₂ O (0.1% TFA)to 60% CH₃ CN/H₂ O (0.1% TFA) and the appropriate product fractions arelyophilized to provide the title compound as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.29 (bs, 2H), 9.08 (bs, 2H), 8.55 (d, 1H),8.33 (d, 1H), 7.93 (d, 1H), 7.68 (m, 2H), 7.57 (m, 3H), 7.20 (d, 1H),7.01 (bs, 1H), 4.45 (Ab, 2H), 4.22 (m, 1H), 3.15 (m, 2H), 2.15 (m, 1H),1.68 (m, 1H). FAB MS, M+H!⁺ =472.

EXAMPLE 43 3-{3-(S)-(5-(2-Amino-pyrimidin-4-yl)-thiophene-2-sulfonyl!-methylamino)-2-oxo-pyrrolidin-1-ylmethyl}-benzamidineditrifluoroacetate

A. 5-(2-Methylsulfinyl-pyrimidin-4-yl)-thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-methylamide and5-(2-Methylsulfonyl-pyrimidin-4-yl)-thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-methylamide

The title compounds are prepared as described in EXAMPLE 28, Part Ausing the mixture of5-(2-methylsulfinyl-pyrimidin-4-yl)-thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide and5-(2-methylsulfonyl-pyrimidin-4-yl)-thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide in place of5-chlorothieno 3,2-b!pyridine-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide. The mixture of crudeproducts are obtained as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.94 (d, 1H), 7.90 (d, 1H), 7.84 (d, 1H), 7.73(m, 1H), 7.62 (m, 1H), 7.46 (m, 3H), 4.93 (m, 1H), 4.43 (AB, 2H), 3.43(s, 3H), 3.27 (m, 2H), 2.97 (s, 3H), 2.41 (m, 1H), 2.06 (m, 1H). FAB MS,M+H!⁺ =516, 532.

B. 3-{3-(S)-(5-(2-Amino-pyrimidin-4-yl)-thiophene-2-sulfonyl!-methylamino-2-oxo-pyrrolidin-1-ylmethyl}-benzamidineditrifluoroacetate

The title compound is prepared as described in EXAMPLE 42 using amixture of 5-(2-methylsulfinyl-pyrimidin-4-yl)-thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-methylamide and5-(2-methylsulfonyl-pyrimidin-4-yl)-thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-methylamide in place of themixture of 5-(2-methylsulfinyl-pyrimidin-4-yl)-thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide and5-(2-methylsulfonyl-pyrimidin-4-yl)-thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide. The crude product ispurified by RP-HPLC eluting in a gradient of 10% CH₃ CN/H₂ O (0.1% TFA)to 60% CH₃ CN/H₂ O (0.1% TFA) and the appropriate product fractions arelyophilized to afford the title compound as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.29 (bs, 2H), 9.03 (bs, 2H), 8.36 (d, 1H),7.98 (d, 1H), 7.75 (d, 1H), 7.68 (m, 1H), 7.56 (m, 3H), 7.21 (d, 1H),6.95 (bs, 1H), 4.88 (m, 1H), 4.44 (AB, 2H), 3.18 (m, 2H), 2.74 (s, 3H),2.07 (m, 1H), 1.89 (m, 1H). IS MS, M+H!⁺ =486.

EXAMPLE 44 3- 3-(S)-(5'-Chloro-2,2'!-bithiophenyl-5-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 5-Chloro- 2,2'!bithiophenyl

The title compound is prepared from 2-chloro-thiophene according to theprocedure described in Bull. Chem. Soc. Japan, 1979, 1126, The crudeproduct is purified by column chromatography eluting with a gradient of5% EtOAc/hexanes to 10% EtOAc/hexanes to afford a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ7.24 (m, 1H), 7.11 (d, 1H), 7.03 (dd, 1H),6.94 (d, 1H), 6.83 (d, 1H). EI MS, M!⁺ =200, 202, Cl pattern.

B. 5'-Chloro- 2,2'!bithiophenyl-5-sulfonyl chloride

The title compound is prepared as described in EXAMPLE 1, Part D using5-chloro- 2,2'!bithiophenyl in place of thianaphthalene. The crudeproduct is purified by column chromatography eluting with a gradient of5% EtOAc/hexanes to 10% EtOAc/hexanes to give a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ7.76 (d, 1H), 7.14 (d, 1H), 7.09 (d, 1H), 6.92(d, 1H), EI MS, M!⁺ =298, 300, Cl pattern.

C. 5'-Chloro- 2,2'!-bithiophenyl-5-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared from3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)-benzonitrile hydrochlorideas described in EXAMPLE 1, Part E using 5'-chloro-2,2'!bithiophenyl-5-sulfonyl chloride in place of benzob!thiophene-2-sulfonyl chloride. The crude product is triturated withEtOAc/CH₂ Cl₂ to yield a beige solid.

¹ H NMR (CDCl₃, 300 MHz) δ7.62 (m, 1H), 7.56 (d, 1H), 7.50 (m, 2H), 7.48(d, 1H), 7.06 (d, 1H), 7.04 (d, 1H), 6.90 (d, 1H), 5.54 (s, 1H), 4.48(s, 2H), 3.93 (m, 1H), 3.27 (m, 2H), 2.65 (m, 1H), 2.15 (m, 1H).

D. 3- 3-(S)-(5'-Chloro-2,2'!-bithiophenyl-5-sulfonylamino-2-oxo-pyrrolidin-1ylmethyl!-benzamidinetrifluoroacetate

5'-Chloro- 2,2'!-bithiophenyl-5-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide is converted to thetitle compound as described in EXAMPLE 1, Part F. The crude product ispurified by RP-HPLC eluting in a gradient of 10% CH₃ CN/H₂ O (0.1% TFA)to 80% CH₃ CN/H₂ O (0.1% TFA) and the appropriate product fractions arelyophilized to provide the title compound as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.30 (bs, 2H), 9.19 (bs, 2H), 8.56 (d, 1H),7.68 (m, 1H), 7.62 (d, 1H), 7.58 (m, 3H), 7.36 (d, 1H), 7.34 (d, 1H),7.19 (d, 1H), 4.46 (AB, 2H), 4.22 (m, 1H), 3.17 (m, 2H), 2.18 (m, 1H),1.70 (m, 1H). FAB MS, M+H!⁺ =495, 497, Cl pattern. Elemental analysiscalculated with 0.9 mol H₂ O: C=42.23%, H=3.52%, N=8.95%, found:C=42.22%, H=3.02%, N=8.62%.

EXAMPLE 45 4-Amino-3- 3-(S)-benzob!thiophene-2-sulfonylamino)-2-oxopyrrolidin-1-ylmethyl!benzamidinetrifluoroacetate

A. Benzo b!thiophene-2-sulfonic acid1-(4-amino-3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!amide

The title compound (0.131 g, 0.307 mmol) is prepared as in EXAMPLE 1,Part E from4-amino-3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)benzonitriledihydrochloride (0.135 g, 0.445 mmol) and benzo b!thiophene-2-sulfonylchloride (0.114 g, 0.49 mmol) in acetonitrile.

¹ H NMR (CDCl₃, 300 MHz) δ7.88-7.94 (m, 3H), 7.48 (m, 2H), 7.36 (dd,1H), 7.28 (d, 1H), 6.58 (d, 1 H), 5.36 (d, 1H), 4.88 (bs, 2H), 4.30 (AB,2H), 3.94 (m, 1H), 3.26 (m, 2H), 2.65 (m, 1H), 2.10 (m, 1H); FAB MS,M+H!⁺ =427.

B. 4-Amino-3- 3-(S)-benzob!thiophene-2-sulfonylamino-2-oxopyrrolidin-1-ylmethyl!benzamidinetrifluoroacetate

The title compound was prepared from benzo b!thiophene-2-sulfonic acid1-(4-amino-3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!amide (0.131 g,0.307 mmol) as described in EXAMPLE 1, Part F. The crude product ispurified by RP-HPLC eluting with a gradient of 10% CH₃ CN/H₂ O (0.1%TFA) to 100% CH₃ CN. The appropriate fractions are lyophilized toprovide 4-amino-3- 3-(S)-benzob!thiophene-2-sulfonylamino-2-oxopyrrolidin-1-ylmethyl!benzamidinetrifluoroacetate as a white solid (0.08 g, 0.14 mmol). ¹ H NMR (DMSO-d₆,300 MHz) δ8.80 (bs, 2H), 8.67 (d, 1H), 8.39 (bs, 2H), 8.0-8.12 (m, 3H),7.47-7.57 (m, 3H), 7.43 (m, 1H), 6.73 (d, .21 (bs, 2H), 4.28 (m, 1H),4.22 (m, 2H), 3.18 (m, 2H), 2.17 (m, 1H), 1.67 (m, 1H). FAB MS, M+H!⁺=444. Elemental analysis calculated with 1.6 mol H₂ O cal. C=45.06%,H=4.33%, N=11.94%, found: C=45.41%, H=4.07%, N=11.47%.

EXAMPLE 46 4-Amino-3- 6-chlorobenzob!thiophene-2-sulfonylamino-2-oxopyrrolidin-1-ylmethyl!benzamidinetrifluoroacetate

A. 6-Chlorobenzo b!thiophene-2-sulfonic acid1-(4-amino-3-cyanobenzyl)-2-oxopyrrolidin-3-(S)-yl!amide

The title compound (0.133 g, 0.288 mmol) is prepared as in EXAMPLE 1,Part E from4-amino-3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)benzonitrile (0.175g, 0.758 mmol) and 6-chlorobenzo b!thiophene-2-sulfonyl chloride (0.203g, 0.758 mmol).

¹ H NMR (CDCl₃, 300 MHz) δ7.89 (s, 1H), 7.86 (d, 1H), 7.82 (d, 1H), 7.45(dd, 1H), 7.37 (dd, 1H), 7.29 (d, 1H), 6.60 (d, 1H), 5.41 (bs, 1H), 4.89(bs, 2H), 4.28 (AB, 2H), 3.97 (m, 1H), 3.28 (m, 2H), 2.64 (m, 1H), 2.09(m, 1H). FAB MS, M+H!⁺ =461,463 Cl pattern.

B. 4-Amino-3- 6-chlorobenzob!thiophene-2-sulfonylamino)-2-oxopyrrolidin-1-ylmethyl!benzamidinetrifluoroacetate

The title compound was prepared from 6-chlorobenzob!thiophene-2-sulfonic acid1-(4-amino-3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!amide (0.131 g,0.284 mmol) as described in EXAMPLE 1, Part F. The crude product ispurified by RP-HPLC eluting with a gradient of 10% CH₃ CN/H₂ O (0.1%TFA) to 100% CH₃ CN. The appropriate fractions are lyophilized toprovide the title compound as a solid (0.126 g, 0.207 mmol).

¹ H NMR (DMSO-d₆, 300 MHz) δ8.80 (bs, 2H), 8.73 (d, 1H), 8.43 (bs, 2H),8.29 (s, 1H), 8.05 (s, 1H), 8.02 (d, 1H), 7.57 (m, 2H), 7.44 (s, 1H),6.73 (dd, 1H), 6.20 (bs, 2H), 4.1-4.25 (m, 3H), 3.20 (m, 2H), 2.18 (m,1H), 1.68 (m, 1H). FAB MS, M+H!⁺ =478, 480, Cl pattern. Elementalanalysis calculated with 1.4 mol H₂ O: C=42.84%, H=3.88%, N=11.35%,found: C=42.84%, H=3.74%, N=11.05%.

EXAMPLE 47 4-Amino-3- 6-chlorobenzob!thiophene-2-sulfonylamino)-2-oxopyrrolidin-1-ylmethyl!hydroxybenzamidinetrifluoroacetate

6-Chlorobenzo b!thiophene-2-sulfonic acid1-(4-amino-3-cyanobenzyl)-2-oxopyrrolidin-3-(S)-yl!amide (0.20 g, 0.434mmol) was converted to the title compound (0.178 g, 0.36 mmol) by themethods described in EXAMPLE 27.

¹ H NMR (DMSO-d₆, 300 MHz) δ12.27 (bs, 1H), 10.80 (bs, 1H), 8.98 (bs,1H), 8.74, (d, 1H), 8.65, (bs, 2H), 8.28, (s, 1H), 8.07 (s, 1H), 8.03(d, 1 H), 7.56 (dd, 1H), 7.39 (dd, 1H), 7.80 (s, 1H), 6.78 (d, 1H), 6.08(bs, 1H), 4.12-4.28 (m, 3H), 3.18 (m, 2H), 2.17 (m, 1H), 1.68 (m, 1H).;Ion spray MS, M!⁺ =495.

EXAMPLE 48 3-2-Oxo-3-(S)-(5-pyridin-3-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 2-Thiophene boronic acid

To a solution of 2-bromothiophene (5.94 mL, 61.3 mmol) in 100 mL ofether at -60° C. is added n-butyl lithium (40 mL of a 1.6M solution inhexanes, 64.4 mmol). After stirring for 30 min, tributylborate (23.7 mL,85.9 mmol) is added and the mixture is stirred for 3 h. The reactionmixture is warmed to room temperature overnight, then treated with 150mL of 1N HCl and washed with ether (2×150 mL). The combined organiclayers are extracted with 1N NaOH. The pH of the aqueous layer isadjusted to <7 using 1N HCl during which time a precipitate forms. Theprecipitate is collected and washed with hexane to give the product asan off-white solid (3.95 g, 30.9 mmol).

¹ H NMR (CDCl₃, 300 MHz) δ7.64 (dd, 1H), 7.59 (dd, 1H), 7.21 (dd, 1H),4.75 (s, 2H). EI, M!⁺ =128.

B. 3-Thiophene-2-yl-pyridine

A mixture of 3-bromopyridine (1.30 mL, 13.5 mmol) andtetrakis(triphenylphosphine) (0.468 g, 0.41 mmol) in 40 mL ofdimethoxyethane is stirred under nitrogen at room temperature for 10min. 2-Thiophene boronic acid (1.90 g, 14.8 mmol) and 20 mL of 1N sodiumcarbonate are added and the resulting mixture is refluxed overnight. Thesolution is cooled to room temperature and filtered through Celite. Thefiltrated is extracted with ether (2×30 mL). The combined organic layersare dried over MgSO₄, filtered and concentrated. The crude product ispurified by column chromatography eluting with a gradient of 10%EtOAc/hexanes to 20% EtOAc/hexanes to afford the title compound (0.355g, 2.20 mmol) as a light yellow oil.

¹ H NMR (CDCl₃, 300 MHz) δ8.89 (d, 1H), 8.52 (dd, 1H), 7.87 (ddd, 1H),7.38 (s, 1H), 7.36 (d, 1H), 7.31, (m, 1H), 7.12 (dd, 1H). EI, M!⁺ =161.

C. 5-Pyridin-3-yl-thiophene-2-sulfonyl choride

To a solution of 3-thiophen-2-yl-pyridine (0.355 g, 2.20 mmol) in 15 mLof THF at -78° C. is added n-BuLi (1.44 mL of a 1.6M solution inhexanes, 2.31 mmol). After stirring for 15 min, SO₂ gas is bubbledthrough the solution for 30 min. The solution is then allowed to warm toroom temperature and stirred overnight. The solution is concentrated todryness and the resulting solid is suspended in 20 mL of hexane.Sulfuryl chloride (0.185 mL, 2.31 mmol) is added and the reactionmixture is stirred for 30 min then a small amount of methylene chlorideis added and the mixture is stirred for another 30 min. The mixture isthen concentrated to dryness and diluted with ethyl acetate and washedwith saturated NaHCO₃ (aq), water and brine. The organic layer is driedover MgSO₄, filtered and concentrated to give a brown solid as the titleproduct (0.452 g, 1.74 mmol) which is used in the subsequent stepwithout further purification.

¹ H NMR (CDCl₃, 300 MHz) δ8.95 (bs, 1H), 8.70 (bs, 1H), 7.95 (d, 1H),7.90 (d, 1H), 7.45 (bs, 1H), 7.44 (d, 1H). EI, M!⁺ =259, 261, Clpattern.

D. 5-Pyridin-3-yl-thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared in CH₃ CN instead of CH₂ Cl₂ as describedin EXAMPLE 1, Part E using 5-pyridin-3-yl-thiophene-2-sulfonyl choridein place of benzo b!thiophene-2-sulfonyl chloride. The crude product isobtained by diluting with ethyl acetate and washing with saturatedsodium bicarbonate (aq), water and brine. The organic layer is driedover MgSO₄, filtered and concentrated. The title product is purified bycolumn chromatography eluting with 5% MeOH/CH₂ Cl₂ to give an off-whitesolid.

¹ H NMR (CDCl₃, 300 MHz) δ8.90 (d, 1H), 8.63 (d, 1H), 7.85 (dd, 1H),7.68 (d, 1H), 7.61 (m, 1H), 7.41-7.45 (m, 3H), 7.34 (dd, 1H), 7.31 (d,1H), 5.45 (s, 1H), 4.50 (s, 2H), 3.95 (m, 1H), 3.25 (m, 2H), 2.68 (m,1H), 2.15 (m, 1H). FAB MS, M+H!⁺ =439.

E. 3-2-Oxo-3-(S)-(8-pyridin-8-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using5-pyridin-3-yl-thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-yl!-amide as the startingmaterial. The crude product is purified by RP-HPLC eluting with agradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.30 (s, 2H), 9.05 (s, 2H), 8.95 (s, 1H),8.63 (d, 1H), 8.55 (d, 1H), 8.16 (d, 1H), 7.65-7.71 (m, 3H), 7.48-7.60(m, 4H), 4.48 (AB, 2H), 4.28 (m, 1H), 3.15 (m, 2H), 2.20 (m, 1H), 1.71(m, 1H). FAB MS, M+H!⁺ =456. Elemental analysis calculated with 0.8 moleof H₂ O: C=43.02%, H=3.55%, N=10.03%; found: C=43.10%, H=3.85%, N=9.98%.

EXAMPLE 49 4-Amino-3-2-oxo-3-(S)-(5-pyridin-3-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 5-Pyridin-3-yl-thiophene-2-sulfonic acid1-(2-amino-5-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared in CH₃ CN instead of CH₂ Cl₂ as describedin EXAMPLE 1, Part E using 5-pyridin-3-yl-thiophene-2-sulfonyl choridein place of benzo b!thiophene-2-sulfonyl chloride and substituting4-amino- 3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)!-benzonitriledihydrochloride for3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)-benzonitrile hydrochloride.The crude product is obtained by diluting with ethyl acetate and washingwith saturated sodium bicarbonate (aq), water and brine. The organiclayer is dried over MgSO₄, filtered and concentrated. The crude productis purified by column chromatography eluting with 50% EtOAc/CH₂ Cl₂ togive a light brown solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.85 (d, 1H), 8.61 (dd, 1H), 7.82 (m, 1H),7.68 (d, 1H), 7.26-7.38 (m, 4H), 6.60 (d, 1H), 5.35 (d, 1H), 4.89 (s,2H), 4.30 (AB, 2H), 3.97 (m, 1H), 3.30 (m, 2H), 2.65 (m, 1H), 2.09 (m,1H). FAB MS, M+H!⁺ =454.

B. 4Amino-3-2-oxo-3-(S)-(5-pyridin-3-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using5-pyridin-3-yl-thiophene-2-sulfonic acid1-(2-amino-5-cyano-benzyl)-2-oxo-pyrrolidin-3-yl!-amide as the startingmaterial. The crude product is purified by RP-HPLC eluting with agradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ8.95 (d, 1H), 8.75 (s, 2H), 8.58 (d, 1H),8.50 (d, 1H), 8.48 (s, 2H), 8.10 (d, 1H), 7.70 (s, 2H), 7.50 (m, 2H),7.40 (d, 1H), 6.71 (d, 1H), 6.20 (bs, 2H), 4.11-4.25 (m, 3H), 3.15 (m,2H), 2.18 (m, 1H), 1.62 (m, 1H). FAB MS, M+H!⁺ =471.

4-Hydroxy-3-2-oxo-3-(S)-(5-pyridin-3-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 5-Pyridin-3-yl-thiophene-2-sulfonic acid {1-5-cyano-2-hydroxy-benzyl!-2-oxo-pyrrolidin-3-(S)-amide

The title compound is prepared as described in EXAMPLE 17, Part G using5-pyridin-3-yl-thiophene-2-sulfonyl choride in place of4,6-dichlorobenzo b!thiophene-2-sulfonyl chloride. The solution isconcentrated, then purified by column chromatography eluting with 10%MeOH/CH₂ Cl₂ to give a brown oil.

¹ H NMR (CDCl₃, 300 MHz) δ8.82 (bs, 1H), 8.61 (bs, 1H), 7.85 (d, 1H),7.65 (d, 1H), 7.63 (d, 1H), 7.55 (s, 1H), 7.53 (dd, 1H), 7.41 (d, 1H),7.31 (m, 1H), 7.28 (d, 1H), 6.91 (d, 1H), 4.41 (s, 2H), 4.12 (m, 1H),3.35 (m, 2H), 2.55 (m, 1H), 2.09 (m, 1H). FAB MS, M+H!⁺ =455.

B. 4Hydroxy-3-2-oxo-3-(S)-(5pyridin-3yl-thiophene-2-sulfonylamino)-pyrrolidin-1ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using5-pyridin-3-yl-thiophene-2-sulfonic acid {1-5-cyano-2-hydroxy-benzyl!-2-oxo-pyrrolidin-3-yl}-amide as the startingmaterial. The crude product is purified by RP-HPLC eluting with agradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ10.91 (s, 1H), 9.06 (s, 2H), 8.98 (s, 1H),8.67 (s, 2H), 8.61 (d, 1H), 8.55 (d, 1H), 8.16 (d, 1H), 7.71 (m, 2H),7.60 (d, 1H), 7.51 (m, 1H), 7.42 (s, 1H), 7.00 (d, 1H), 4.35 (AB, 2H),4.21 (m, 1H), 3.21 (m, 2H), 2.20 (m, 1H), 1.71 (m, 1H). FAB MS, M+H!⁺=472.

EXAMPLE 51 4-Hydroxy-3-2-oxo-3-(S)-(5-pyridin-N-oxide-3-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 5-Pyridin-3-yl-thiophene-2-sulfonic acid {1-2-(tert-butyl-dimethyl-silanyloxy)-5-cyano-benzyl!-2-oxo-pyrrolidin-3-(S)-yl}amide

Imidazole (0.094 g, 1.37 mmol) and t-butyldimethylchlorosilane (0.099 g,0.66 mmol) are added to a solution of5-pyridin-3-yl-thiophene-2-sulfonic acid {1-5-cyano-2-hydroxy-benzyl!-2-oxo-pyrrolidin-3-yl}-amide (0.25 g, 0.55mmol) in 10 mL of DMF. The resulting mixture is stirred overnight, thendiluted with EtOAc and washed with saturated NaHCO₃ (aq) and brine. Theorganic layer is dried over MgSO₄, filtered and concentrated. Theproduct is purified by column chromatography eluting with 3% MeOH/CH₂Cl₂ to give a brown oil (0.236 g, 0.41 mmol).

1H NMR (CDCl₃, 300 MHz) δ8.90 (bs, 1H), 8.52 (bs, 1H), 7.79 (m, 1H),7.60 (d, 1H), 7.56 (dd, 1H), 7.45 (d, 1H), 7.39 (m, 1H), 7.28 (d, 1H),7.22 (d, 1H), 6.80 (d, 1H), 6.70 (s, 1H), 4.40 (s, 2H), 4.03 (m, 1H),3.21 (m, 2H), 2.60 (m, 1H), 2.10 (m, 1H), 0.92 (s, 9H), 0.20 (s, 6H).

B. 5-Pyridin-N-oxide-3-yl-thiophene-2-sulfonic acid {1-2-(tert-butyl-dimethyl-silanyloxy)-5-cyano-benzyl!-2-oxo-pyrrolidin-3-(S)-yl}-amide

To a solution of 5-pyridin-3-yl-thiophene-2-sulfonic acid {1-2-(tert-butyl-dimethyl-silanyloxy)-5-cyano-benzyl!-2-oxo-pyrrolidin-3-(S)-yl}-amide(0.18 g, 0.32 mmol) in 10 mL of CH₂ Cl₂, is added m-chloroperbenzoicacid (0.17 g, 0.63 mmol). The mixture is stirred overnight then dilutedwith CH₂ Cl₂ and washed with saturated NaHCO₃ (aq) and brine. Theorganic layer is dried over MgSO4, filtered and concentrated to give thetitle product as a yellow oil (0.18 g, 0.32 mmol) which is used in thesubsequent step without further purification.

¹ H NMR (CDCl₃, 300 MHz) δ10.70 (bs, 1H), 8.85 (s, 1H), 8.42 (d, 1H),8.00 (s, 1H), 7.95 (d, 1H), 7.85 (s, 1H), 7.80 (d, 1H), 7.70 (m, 1H),7.35 (d, 1H), 6.90 (d, 1H), 6.82 (d, 1H), 4.44 (s, 2H), 4.18 (m, 1H),3.31 (m, 2H), 2.62 (m, 1H), 2.20 (m, 1H), 0.92 (s, 9H), 0.25 (s, 6H).FAB MS, M+H!⁺ =585.

C. 4-Hydroxy-3-2-oxo-3-(S)-(5-pyridin-N-oxide-3-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using5-pyridin-N-oxide-3-yl-thiophene-2-sulfonic acid {1-2-(tert-butyl-dimethyl-silanyloxy)-5-cyano-benzyl!-2-oxo-pyrrolidin-3-(S)-yl}-amide.The crude product is purified by RP-HPLC eluting with a gradient of 10%CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA). The appropriatefractions are lyophilized to provide the title compound as a whitesolid.

¹ H NMR (DMSO-d₆, 300 MHz) δ10.91 (s, 1H), 9.05 (s, 2H), 8.75 (s, 1H),8.65 (s, 2H), 8.60 (d, 1H), 8.21 (d, 1H), 7.76 (d, 1H), 7.70 (d, 1H),7.66 (d, 1H), 7.60 (dd, 1H), 7.49 (m, 1H), 7.41 (d, 1H), 6.98 (d, 1H),4.35 (AB, 2H), 4.19 (m, 1H), 3.21 (m, 2H), 2.25 (m, 1H), 1.75 (m, 1H).FAB MS, M+H!⁺ =488.

EXAMPLE 52 3-2-Oxo-3-(S)-(5-pyridin-4-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 4-Thiophen-2-yl-pyridine

The title compound is prepared as described in EXAMPLE 48, Part B using4-bromopyridine in place of 3-bromopyridine. The crude product ispurified by column chromatography eluting with a gradient of 20%EtOAc/hexanes to 40% EtOAc/hexanes to afford the title product as awhite solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.60 (d, 2H), 7.51 (m, 1H), 7.49 (d, 2H), 7.42(dd, 1H), 7.14 (dd, 1H). EI, M!⁺ =161.

B. 5-Pyridin-4-yl-thiophene-2-sulfonyl choride

The title compound is prepared as described in EXAMPLE 48, Part C using4-thiophen-2-yl-pyridine in place of 3-thiophen-2-yl-pyridine. The titlecompound is used in the subsequent step without further purification.

¹ H NMR (CDCl₃, 300 MHz) δ8.75 (d, 2H), 8.60 (d, 1H), 7.90 (d, 1H), 7.51(d, 2H). EI, M!⁺ =259, 261, Cl pattern.

C. 5-Pyridin-4-yl-thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared in CH₃ CN instead of CH₂ Cl₂ as describedin EXAMPLE 1, Part E using 5-pyridin-4-yl-thiophene-2-sulfonyl choridein place of benzo b!thiophene-2-sulfonyl chloride. The crude product isobtained by diluting with ethyl acetate and washing with saturatedsodium bicarbonate (aq), water and brine. The organic layer is driedover MgSO₄, filtered and concentrated. The title product is purified bycolumn chromatography eluting with 5% MeOH/CH₂ Cl₂ to give a light brownsolid.

¹ H NMR (CDCl₃, 300 MHz) δ8.70 (d, 2H), 7.68 (d, 1H), 7.60 (m, 1H),7.40-7.50 (m, 6H), 5.65 (bs, 1H), 4.48 (AB, 2H), 3.98 (m, 1H), 3.28 (m,2H), 2.68 (m, 1H), 2.17 (m, 1H). FAB MS, M+H!⁺ =439.

D. 3-2-Oxo-3-(S)-(5-pyridin-4-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using5-pyridin-4-yl-thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-yl!-amide as the startingmaterial. The crude product is purified by RP-HPLC eluting with agradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.30 (s, 2H), 9.05 (s, 2H), 8.69 (d, 2H),8.67 (d, 1H), 7.89 (d, 1H), 7.85 (d, 2H), 7.77 (d, 1H), 7.68 (m, 2H),7.51-7.60 (m, 3H), 4.42 (AB, 2H), 4.26 (m, 1H), 3.18 (m, 2H), 2.21 (m,1H), 1.70 (m, 1H). FAB MS, M+H!⁺ =456.

EXAMPLE 53 3-3-(S)-(4-Chloro-thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 4-Chloro-5-trimethylsilanyl-thiophene-2-sulfonyl chloride

The title compound is prepared as described in EXAMPLE 48, Part C using(3-chloro-thiophen-2-yl)-trimethylsilane (prepared using the methoddescribed in WO 94/12505, PCT/US93/08613) in place of3-thiophen-2-yl-pyridine.

¹ H NMR (CDCl₃, 300 MHz) δ7.10 (s, 1H), 0.30 (s, 9H). EI, M!⁺ =288, 290,Cl pattern.

B. 4-Chloro-5-trimethylsilanyl-thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide and4-chloro-thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared in CH₃ CN instead of CH₂ Cl₂ as describedin EXAMPLE 1, Part E using4-chloro-5-trimethylsilanyl-thiophene-2-sulfonyl choride in place ofbenzo b!thiophene-2-sulfonyl chloride. The crude product is obtained bydiluting with ethyl acetate and washing with saturated sodiumbicarbonate (aq), water and brine. The organic layer is dried overMgSO₄, filtered and concentrated. The title product is purified bycolumn chromatography eluting with 3% MeOH/CH₂ Cl₂ to give two products.4-chloro-5-trimethylsilanyl!-thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

¹ H NMR (CDCl₃, 300 MHz) δ7.60 (m, 1H), 7.45-7.50 (m, 3H), 7.30 (s, 1H),5.80 (bs, 1H), 4.50 (AB, 2H), 3.85 (m, 1H), 3.28 (m, 2H), 2.62 (m, 1H),2.15 (m, 1H), 0.35 (s, 9H), FAB MS, M+H!⁺ =396, 398, Cl pattern.

4-chloro-thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

¹ H NMR (CDCl₃, 300 MHz) δ7.69 (m, 1H), 7.56 (d, 1H), 7.50 (m, 3H), 7.08(d, 1H), 5.83 (bs, 1H), 4.48 (s, 2H), 3.85 (s, 1H), 3.21 (m, 2H), 2.60(m, 1H), 2.11 (m, 1H), FAB MS, M+H!⁺ =468, 470, Cl pattern.

C. 3-3-(S)-(4-Chloro-thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using4-chloro-thiophene-2-sulfonic acid1-(3-cyano-benzyl)-2-oxo-pyrrolidin-3-yl!-amide as the startingmaterial. The crude product is purified by RP-HPLC eluting with agradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.25 (bs, 2H), 8.91 (bs, 2H), 8.65 (m, 1H),7.95 (d, 1H), 7.68 (m, 1H), 7.55 (m, 2H), 7.22 (d, 1H), 4.41 (AB, 2H),4.20 (m, 1H), 3.18 (m, 2H), 2.10 (m, 1H), 1.75 (m, 1H). MS, M+H!⁺ =413,415, Cl pattern.

EXAMPLE 54 3-{3-(S)-5-(5-Chloropyridin-3-yl)-thiophene-2-sulfonylamino!-2-oxopyrrolidin-1-ylmethyl}benzamidinetrifluoroacetate

A. 5-Chloro-3-(thiophene-2-yl)pyridine

The title compound (0.21 g, 1.08 mmol) was prepared from3,5-dichloropyridine (1.0 g, 6.76 mmol) and 2-thiophene boronic acid(0.95 g, 7.43 mmol) as described in EXAMPLE 48, Part B.

¹ H NMR (CDCl₃, 300 MHz) δ8.73 (d, 1H), 8.47 (d, 1H), 7.83 (dd, 1H),7.34 (two dd, 2H), 7.10 (dd, 1H).

B. 5-(5-Chloropyridin-3-yl}thiophene-2-sulfonylchloride

5-(5-Chloropyridin-3-yl)thiophene-2-sulfonylchloride (0.21 g, 1.08 mmol)was prepared as described in EXAMPLE 48, Part C from5-chloro-3-(thiophene-2-yl)pyridine.

¹ H NMR (CDCl₃, 300 MHz) δ8.88 (d, 1H), 8.66 (d, 1H), 7.94 (m, 1H), 7.65(AB, 2H); EI MS, M!⁺ =293,295.

C. 5-(5-Chloropyridin-3-yl)thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!amide

The title compound (0.012 g, 0.288 mmol) is prepared as in EXAMPLE 1,Part E from 3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)benzonitrile(0.039 g, 0.153 mmol) and 6-chlorobenzo b!thiophene-2-sulfonyl chloride(0.045 g, 0.153 mmol) in acetonitrile.

¹ H NMR (CDCl₃, 300 MHz) δ8.74 (d, 1H), 8.58 (d, 1H), 7.88 (dd, 1H),7.61 (m, 1H), 7.52 (AB, 2H), 7.47 (m, 3H), 5.64 (bs, 1 H), 4.47(AB, 2H)3.97 (dd, 3H) 3.27 (m, 2H), 2.64 (m, 1H), 2.16 (m, 1H); EI MS, M!⁺ =473,475, Cl pattern.

D. 3-{3-(S)-5-(5-Chloropyridin-3-yl)-thiophene-2-sulfonylamino!-2-oxo-pyrrolidin-1ylmethyl}benzamidinetrifluoroacetate

The title compound was prepared from5-(5-chloropyridin-3-yl)thiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!amide (0.012 g, 0.025 mmol)as described in EXAMPLE 1, Part F. The crude product is purified byRP-HPLC eluting with a gradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 100%CH₃ CN. The appropriate fractions are lyophilized to provide 3-{3-(S)-5-(5-chloropyridin-3-yl)-thiophene-2-sulfonylamino!-2-oxopyrrolidin-1-ylmethyl}benzamidinetrifluoroacetate as a white solid (0.005 g, 0.14 mmol). ¹ H NMR(DMSO-d₆, 300 MHz) δ9.28 (bs), 8.89 (bs), 8.60 (m), 8.37 (m), 7.75 (AB),7.67 (m), 7.56 (m4.44 (AB), 4.24 (m, 2H), 3.17 (m), 2.20 (m), 1.69 (m).FAB MS, M+H!⁺ =490, 492, C pattern.

EXAMPLE 55 3-3-(S)-(4-Chloro-5-pyridin-3-ylthiophene-2-sulfonylamino)-2-oxopyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. (3-Chlorothiophene-2-yl)pyridine

3-Chlorothiophene-2-boronic acid (1.5 g, 9.24 mmol), prepared asdescribed in EXAMPLE 48, Part A from 3-chlorothiophene was reacted with3-bromopyridine (0.81 mL, 8.4 mmol) as described in EXAMPLE 48, Part Bto give the title compound (0.232 g, 1.19 mmol).

¹ H NMR (CDCl₃, 300 MHz) δ8.89 (dd, 1H), 8.58 (dd, 1H), 7.98 (ddd, 1H),7.36 (ddd, 1H), 7.19 (AB, 2H).

B. 3-Chloro-5-pyridin-3-ylthiophene-2-sulfonylchloride

(3-Chlorothiophene-2-yl)pyridine (0.232 g, 1.19 mmol) was reacted asdescribed in EXAMPLE 48, Part C to give3-chloro-5-(pyridin-3-yl)thiophene-2-sulfonyl chloride (0.154 g, 0.612mmol) which is contaminated with some butylated material.

¹ H NMR (CDCl₃, 300 MHz) δ8.92 (bs, 1H), 8.87 (d, 1H), 8.05 (dd, 1H),7.84 (s, 1H), 7.47 (dd, 1H); EI MS, M!⁺ =293, 295, Cl pattern.

C. 4-Chloro-5-pyridin-3-ylthiophene-2-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!amide

The title compound (0.063 g, 0.133 mmol) is prepared as in EXAMPLE 1,Part E from 3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)benzonitrile(0.154 g, 0.612 mmol) and3-chloro-5-pyridin-3-ylthiophene-2-sulfonylchloride (0.180 g, 0.612mmol) in acetonitrile.

¹ H NMR (CDCl₃, 300 MHz) 8.86 (bs, 1H), 8.70 (dd, 1H), 7.98 (ddd, 1H),7.42-7.55 (m, 4H), 7.56-7.66 (m,2H), 6.64 (bs, 1H), 4.50 (AB, 2H) 4.08(dd, 3H) 3.28 (m, 2H), 2.67 (m, 1H), 2.18 (m, 1H).

D. 3-3-(S)-(4-Chloro-5-pyridin-3-ylthiophene-2-sulfonylamino)-2-oxopyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

The title compound was prepared from 4-chloro-5-pyridin-3-ylthiophene-2-sulfonic acid 1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!amide(0.063 g, 0.133 mmol) as described in EXAMPLE 1, Part F. The crudeproduct is purified by RP-HPLC eluting with a gradient of 10% CH₃ CN/H₂O (0.1% TFA) to 100% CH₃ CN to give 3-3-(S)-(4-chloro-5-pyridin-3-ylthiophene-2-sulfonylamino)-2-oxopyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate as a white solid (0.076 g, 0.106 mmol). ¹ H NMR(DMSO-d₆, 300 MHz) δ9.32 (bs, 2H), 9.08 (bs, 2H), 8.88 (bs, 1H), 8.82(d, 1H), 8.70 (d, 1H), 8.13 (dd, 1H), 7.87 (s, 1H), 7.72 (m,1H), 7.51(m, 4H4.45 (AB, 2H), 4.31 (m, 1H), 3.21 (m, 2H), 2.33 (m, 1H), 1.79 (m,1H). Ion spray MS, M+H!⁺ =490, 492, Cl pattern.

EXAMPLE 56 4-Hydroxy-3- 3-(S)-(6-chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidetrifluoroacetate

A. 6-Chlorobenzo b!thiophene-2-sulfonic acid1-(5-cyano-2-hydroxy-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared from 3-(3-amino-2-oxo-pyrrolidin-1-yl)-methyl!-4-hydroxy-benzonitrilehydrochloride as described in EXAMPLE 17, Part G substituting6-chlorobenzo b!thiophene-2-sulfonyl chloride in place of4,6-dichlorobenzo b!thiophene-2-sulfonyl chloride. The crude product istriturated with Et₂ O to give the title compound as a white solid.

1H NMR (CDCl₃) δ8.15 (d, 1H), 7.95 (s, 1H), 7.82 (m, 2H), 7.50 (s, 1H),7.40 (m, 2H), 6.94 (d, 1H), 4.46 (d, 1H), 4.35 (d, 1H), 4.15 (t, 1H),3.30 (m, 2H), 2.40 (m, 1H), 1.95 (m, 1H). FAB MS, M+H!⁺ =462, 464, Clpattern.

B. 4Hydroxy-3- 3-(S)-(6chlorobenzob!thiophene-2sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamide

The title compound is prepared as described in EXAMPLE 1, Part F using6-chlorobenzo b!thiophene-2-sulfonic acid1-(5-cyano-2-hydroxy-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide as thestarting material. The crude product is purified by RP-HPLC eluting witha gradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ10.80 (bs, 1H), 9.00 (bs, 2H), 8.82 (d, 1H),8.62 (bs, 2H), 7.98 (m, 2H), 7.90 (d, 1H), 7.47 (m, 2H), 6.65 (d, 1H),4.49 (AB, 2H), 4.32 (m, 1H), 3.32 (m, 2H), 2.30 (m, 1H), 1.82 (m, 1H).FAB MS, M+H!⁺ =479, 481, Cl pattern.

EXAMPLE 57 3-3-(S)-(1-Aminoisoquinoline-6-sulfonylamino)-2-oxopyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate

A. 3-(3-Bromophenyl)acrylic acid

3-Bromobenzaldehyde (10.0 g, 54 mmol), malonic acid (10.1 g, 97 mmol)and piperidine (0.267 mL, 2.7 mmol) are treated with pyridine (30 mL)and heated at 100° C. for 3.5 h. The reaction mixture is cooled to roomtemperature, poured into 200 ml of 20% hydrochloric acid at 0° C.; theresultant precipitate is collected, washed with copious amounts of waterand dried in a vacuum dessicator overnight to afford the title compound(11.64 g, 51 mmol).

¹ H NMR (DMSO-d₆, 300 MHz) δ12.55 (bs, 1H), 7.96 (s, 1H), 7.70 (d, 1H),7.5-7.64 (m, 2H), 7.38 (m, 1H), 6.62 (d, 1H). El MS, M⁺ =226, 228, Brpattern.

B. 6-Bromoisoquinolin-1-one

3-(3-Bromophenyl)acrylic acid (10.0 g, 44 mmol) is subjected methodsdescribed in EXAMPLE 23, Part B to give 5.6 g of a yellow solid. Thismaterial is washed with hot ethanol/ethyl acetate to give the titlecompound as a white solid (2.54 g, 11 mmol).

¹ H NMR (DMSO-d₆, 300 MHz) δ11.32 (bs, 1H), 8.05 (bs, 1H), 7.92 (s, 1H),7.58 (d, 1H), 7.23 (m, 1H), 6.49 (d, 1H). EI MS, M⁺ =223, 225, Brpattern.

C. 6-Bromo-1-Chloroisoquinoline

6-Bromoisoquinolin-1-one (2.54 g, 11 mmol) is converted to the titlecompound (2.69 g, 11 mmol) by the method described in EXAMPLE 23, PartC.

¹ H NMR (CDCl₃, 300 MHz) δ8.30 (d, 1H), 8.19 (d, 1H), 8.04 (s, 1H), 7.78(d, 1H), 7.52 (d, 1H), 7.27 (s, 1H), 6.49 (d, 1H). EI MS, M⁺ =241, 243.

D. 1-Chloroisoquinoline-2-sulfonyl chloride

6-Bromo-1-Chloroisoquinoline (2.69 g, 11 mmol) is converted to the titlecompound by the method described in EXAMPLE 1, Part D except that thecrude product was washed with hexane to give a yellow solid (3.6 g)which was used without further purification; EI MS, M⁺ =261, 263.

E. 1-Chloroisoquinoline-6-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!amide

3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)benzonitrile hydrochloride(1.7 g, 6.8 mmol) is reacted with 1-Chloroisoquinoline-2-sulfonylchloride (3.54 g, 13.6 mmol) as described in EXAMPLE 1, Part E. Thecrude product was purified by flash chromatography (2% MeOH/CH₂ Cl₂) toyield the title compound (1.14 g, 2.58 mmol).

¹ H NMR (CDCl₃, 300 MHz) δ8.49 (m, 2H), 8.40 (d, 1H), 8.12 (dd, 1H),7.74 (d, 1H), 7.57 (m, 1H), 7.45 (d, 1H), 7.39 (d, 1H), 5.83 (bs, 1H),4.44 (AB, 2H), 3.9 (dd, 1H) 3.20 (dd, 2H), 2.61 (m, 1H), 2.08 (m, 1H);FAB MS, M+H!⁺ =441, 443.

F. 1-Aminoisoquinoline-6-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!amide

1-Aminoisoquinoline-6-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!amide (0.20 g, 0.45 mmol)and phenol (3 g) were heated together at 70° C. for 5 min. treated withammonium acetate (2.5 g) and heated to 115° C. for an additional 7 h.The reaction is cooled to room temperature, diluted with ethyl acetateand partitioned with 1N sodium hydroxide. The aqueous layer wassaturated with sodium chloride and washed with fresh ethyl acetate(2×100 mL). The organic layers are combined, dried (sodium sulfate),concentrated and chromatographed (5% MeOH/CH₂ Cl₂) to afford the titlecompound (0.108 g, 0.26 mmol).

¹ H NMR (CDCl₃, 300 MHz) δ8.28 (s, 1H), 8.03 (d, 1H), 7.92 (AB, 2H),7.55 (m, 1H), 7.4-7.48 (m, 3H), 7.08 (d, 1 H), 5.50 (bs, 3H), 4.34 (AB,2H), 3.89 (dd, 1H) 3.22 (m, 2H), 2.58 (m, 1H), 2.09 (m, 1H); FAB MS,M+H!⁺ =422.

G. 3-3-(S)-(1-Aminoisoquinoline-6-sulfonylamino)-2-oxopyrrolidin-1-ylmethyl!-benzamidinebistrifluoroacetate

1-Aminoisoquinoline-6-sulfonic acid1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!amide (0.108 g, 0.26 mmol)is treated as described in EXAMPLE 1, Part F and purified by HPLC toobtain the title compound as a white solid (0.10 g, 0.15 mmol).

¹ H NMR (DMSO-d₆, 300 MHz) δ9.39 (bs, 2H), 9.21 (bs, 4H), 8.69 (d, 1H),8.60 (d, 1H), 8.46 (d, 1H), 8.12 (dd, 1H), 7.80 (d, 1H), 7.67 (d, 1H),7.5-7.6 (m, 3H), 7.38 (d, 1H), 4.39 (bs, 2H), 4.26 (m, 1H) 3.14 (dd,2H), 2.18 (m, 1H), 1.67 (m, 1H); FAB MS, M+H!⁺ =439. Elemental analysiscalculated with 2 mol H₂ O: C=42.09%, H=4.13%, N=11.78%, found:C=42.17%, H=3.90%, N=11.38%.

EXAMPLE 58 4-Fluoro-3-3-(S)-(5-pyridin-3-yl-thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidetrifluoroacetate

A. 5-Bromo-2-fluoro-benzyl alcohol

To a solution of 5-bromo-2-fluoro-benzaldehyde (6.10 g, 30.0 mmol) in 30mL of THF at 0° C. is added 5 mL of sodium borohydride (2.0M solution intriglyme, 10.0 mmol). The reaction mixture is stirred at 0° C. for 25min and then quenched by the addition of 1N HCl. The mixture is dilutedwith EtOAc and the layers are separated. The organic layer is washedwith H₂ O and saturated NaCl. The organic layer is dried over MgSO₄,filtered and concentrated. The crude product is purified by columnchromatography eluting with 15% EtOAc/hexanes to afford the titlecompound (6.00 g, 29.3 mmol).

¹ H NMR (CDCl₃, 300 MHz) δ7.55 (dd, 1H), 7.36 (m, 1H), 6.89 (t, 1H),4.71 (d, 2H), 2.11 (bs, 1H). EI MS, M!⁺ =204, 206, Br pattern.

B. 5-Bromo-2-fluoro-benzyl bromide

To a solution of 5-bromo-2-fluoro-benzyl alcohol (3.10 g, 15.1 mmol) in30 mL of THF at 10° C. is added triphenyl phosphine (4.10 g, 15.6 mmol)followed by N-bromosuccinimide (2.67 g, 15.0 mmol). The ice bath isremoved and the resulting solution is stirred for 20 min at roomtemperature. The crude product is purified by column chromatographyeluting with 5% EtOAc/hexanes to give the title compound (3.90 g, 14.5mmol).

¹ H NMR (CDCl₃, 300 MHz) δ7.50 (dd, 1H), 7.37 (m, 1H), 6.92 (t, 1H),4.42 (s, 2H). EI MS, M!⁺ =266, 268, 270; 2 Br pattern.

C.3-(S)-(tert-Butoxy-carbonyl-amino)-1-(5-bromo-2-fluoro-benzyl)-pyrrolidin-2-one

The title compound is prepared as described in EXAMPLE 1, Part Bsubstituting 5-bromo-2-fluoro-benzyl bromide for α-bromo-m-toluylnitrile. The crude material is purified by column chromatography elutingwith 3:1:1 EtOAc:hexane:CH₂ Cl₂.

¹ H NMR (CDCl₃, 300 MHz) δ7.40 (m, 2H), 6.96 (t, 1H), 5.20 (bs, 1H),4.50 (s, 2H), 4.18 (m, 1H), 3.28 (m, 2H), 2.64 (m, 1H), 1.90 (m, 1H),1.45 (s, 9H). EI MS, M!⁺ =387, 389, Br pattern.

D.3-(S)-(tert-Butoxy-carbonyl-amino)-1-(5cyano-2-fluorobenzyl)-pyrrolidin-2-one

The title compound is prepared as described in EXAMPLE 3, Part Csubstituting3-(S)-(tert-butoxy-carbonyl-amino)-1-(5-bromo-2-fluoro-benzyl)-pyrrolidin-2-onefor (5-iodo-thiophene-3-yl)methanol. The crude material is purified bycolumn chromatography eluting with 3:1:1 EtOAc:hexane:CH₂ Cl₂.

¹ H NMR (CDCl₃, 300 MHz) δ7.70 (bd, 1H), 7.61 (m, 1H), 7.19 (t, 1H),5.18 (bs, 1H), 4.63 (d, 1H), 4.50 (d, 1H), 4.18 (m, 1H), 3.32 (m, 2H),2.64 (m, 1H), 2.00 (m, 1H), 1.47 (s, 9H). EI MS, M!⁺ =334.

E. 3-(S)- (3-Amino-2-oxo-pyrrolidin-1-yl)-methyl!-4-fluoro-benzonitrilehydrochloride

The title compound is prepared as described in Example 1, Part Csubstituting3-(S)-(tert-butoxy-carbonyl-amino)-1-(5-cyano-2-fluoro-benzyl)-pyrrolidin-2-onefor 1-(3-cyanobenzyl)-2-oxo-pyrrolidin-3-(S)-yl!-carbamic acidtert-butyl ester. The title compound is obtained as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ8.73 (bs, 3H), 7.92 (m, 1H), 7.86 (d, 1H),7.50 (t, 1H), 4.54 (s, 2H), 4.10 (m, 1H), 3.38 (m, 2H), 2.42 (m, 1H),2.07 (m, 1H). EI MS, M!⁺ =233.

F. 5-Pyridin-3-yl-thiophene-2-sulfonic acid1-(5-cyano-2-fluoro-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide

The title compound is prepared from 3-(S)-(3-amino-2-oxo-pyrrolidin-1-yl)-methyl!-4-fluoro-benzonitrilehydrochloride as described in EXAMPLE 17, Part G substituting5-pyridin-3-yl-thiophene-2-sulfonyl chloride in place of4,6-dichlorobenzo b!thiophene-2-sulfonyl chloride. The crude product istriturated with Et₂ O to give the title compound as a white solid

¹ H NMR (CDCl₃ +CD₃ OD, 300 MHz) δ8.87 (s, 1H), 8.55 (d, 1H), 8.08 (d,1H), 7.73 (m, 3H), 7.53 (m, 1H), 7.48 (d, 1H), 7.28 (t, 1H), 4.54 (AB,2H), 4.20 (m, 1H), 3.34 (m, 2H), 2.49 (m, 1H), 1.98 (m, 1H). FAB MS,M+H!⁺ =457.

G. 4-Fluoro-3-3-(S)-(5-pyridin-3-yl-thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidetrifluoroacetate

The title compound is prepared as described in EXAMPLE 1, Part F using5-pyridin-3-yl-thiophene-2-sulfonic acid1-(5-cyano-2-fluoro-benzyl)-2-oxo-pyrrolidin-3-(S)-yl!-amide as thestarting material. The crude product is purified by RP-HPLC eluting witha gradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 80% CH₃ CN/H₂ O (0.1% TFA).The appropriate fractions are lyophilized to provide the title compoundas a white solid.

¹ H NMR (CD₃ OD, 300 MHz) δ8.77 (s, 1H), 8.44 (d, 1H), 8.03 (d, 1H),7.67 (m, 2H), 7.58 (d, 1H), 7.45 (d, 1H), 7.41 (m, 1H), 7.28 (t, 1H),5.10 (s, 1H), 4.47 (s, 2H), 4.13 (m, 1H), 3.23 (m, 2H), 2.28 (m, 1H),1.81 (m, 1H). FAB MS, M+H!⁺ =473.

EXAMPLE 59 2-Chloroquinoline-6-sulfonic acid {1-3-(aminoiminomethyl)-benzyl!-2-oxo-pyrrolidin-3-(S)-yl}amidetrifluoroacetate

A. 2-Chloro-6-bromoquinoline

The title compound is prepared from 4-bromoaniline and cinnamoylchloride according to the procedure described in J. Chem. Soc., PerkinTrans. I, 1972, 1648. The crude 6-bromo-1H-quinolin-2-one intermediateobtained is triturated in Et₂ O/hexanes and filtered to give a beigesolid which is used directly in the chlorination step. The crude productis recrystallized in MeOH to afford the title compound as a beige solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.01 (d, 1H), 7.98 (s, 1H), 7.89 (m, 1H), 7.80(m, 1H), 7.40 (d, 1H).

B. 2-Chloroquinoline-6-sulfonyl chloride

The title compound is prepared as described in EXAMPLE 1, Part D using2-chloro-6-bromoquinoline in place of thianaphthalene. The crude productis triturated with hexanes to give a beige solid and is of sufficientpurity to be used in the subsequent step.

¹ H NMR (CDCl₃, 300 MHz) δ8.60 (s, 1H), 8.31 (d, 1H), 8.28 (m, 2H), 7.60(d, 1H).

C. 2-Chloroquinoline-6-sulfonic acid1-(3-cyanobenzyl-2-oxopyrrolidin-3-(S)-yl!-amide

The title compound is prepared from3-(3-(S)-amino-2-oxo-pyrrolidin-1-ylmethyl)-benzonitrile hydrochlorideas described in EXAMPLE 1, Part E using 2-chloroquinoline-6-sulfonylchloride in place of benzo b!thiophene-2-sulfonyl chloride. The productis obtained as a white solid.

¹ H NMR (CDCl₃, 300 MHz) δ8.50 (s, 1H), 8.25 (d, 1H), 8.18 (m, 1H), 8.14(m, 1H), 7.58 (m, 1H), 7.52 (d, 1H), 7.44 (m, 3H), 5.68 (bs, 1H), 4.45(AB, 2H), 3.89 (m, 1H), 3.22 (m, 2H), 2.63 (m, 1H), 2.08 (m, 1H).

D. 2-Chloroquinoline-6-sulfonic acid {1-3-(aminoiminomethyl)-benzyl!-2-oxopyrrolidin-3-(S)-yl}amidetrifluoroacetate

2-Chloroquinoline-6-sulfonic acid1-(3-cyanobenzyl)-2-oxopyrrolidin-3-(S)-yl!-amide is converted to thetitle compound as described in EXAMPLE 1, Part F. The crude product ispurified by RP-HPLC eluting in a gradient of 10% CH₃ CN/H₂ O (0.1% TFA)to 60% CH₃ CN/H₂ O (0.1% TFA) and the appropriate product fractions arelyophilized to provide the title compound as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.29 (bs, 2H), 9.13 (bs, 2H), 8.68 (d, 1H),8.64 (s, 1H), 8.46 (d, 1H), 8.19 (m, 1H), 8.13 (m, 1H), 7.75 (d, 1H),7.66 (m, 1H), 7.55 (m, 3H), 4.42 (AB, 2H), 4.21 (m, 1H), 3.10 (m, 2H),2.07 (m, 1H), 1.62 (m, 1H). IS MS, M+H!⁺ =458, 460, Cl pattern.Elemental analysis calculated with 1.9 mol H₂ O cal. C=45.54%, H=4.13%,N=11.54%, found: C=45.53%, H=3.49%, N=10.79%.

EXAMPLE 60 2-Aminoquinoline-6-sulfonic acid {1-3-(aminoiminomethyl)-benzyl!-2-oxo-pyrrolidin-3-(S)-yl}amideditrifluoroacetate

2-Chloroquinoline-6-sulfonic acid {1-3-(aminoiminomethyl)-benzyl!-2-oxopyrrolidin-3-(S)-yl}amidetrifluoroacetate (0.25 g, 0.50 mmol) and phenol (0.80 g, 8.25 mmol) aremelted together at 80° C. for 5 min. To the mixture is added ammoniumacetate (0.64 g, 8.25 mmol) and heating is continued at 120° C. for 2.5h. At this time, more NH₄ OAc (s) is added. After 1 h, the reactionmixture is cooled to room temperature and partitioned between EtOAc and0.5N HCl. The layers are separated and the aqueous phase is extractedwith EtOAc. The aqueous layer is concentrated in vacuo to a small volume(˜5 mL). The solution of crude product is purified by RP-HPLC eluting ina gradient of 10% CH₃ CN/H₂ O (0.1% TFA) to 40% CH₃ CN/H₂ O (0.1% TFA)and the appropriate product fractions are lyophilized to provide thetitle compound as a white solid.

¹ H NMR (DMSO-d₆, 300 MHz) δ9.29 (bs, 2H), 9.21 (bs, 2H), 9.03 (bs, 2H),8.46 (d, 1H), 8.44 (s, 1H), 8.37 (d, 1H), 8.13 (d, 1H), 7.75 (d, 1H),7.68 (m, 1H), 7.56 (m, 3H), 7.15 (d, 1H), 4.40 (s, 2H), 4.16 (m, 1H),3.10 (m, 2H), 2.07 (m, 1H), 1.62 (m, 1H). IS MS, M+H!⁺ 439. Elementalanalysis calculated with 2.4 mol H₂ O: C=42.33%, H=4.09%, N=11.85%,found: C=42.33%, H=3.69%, N=11.39%.

The molecules described herein inhibit blood coagulation by virtue oftheir ability to inhibit the penultimate enzyme in the coagulationcascade, controlling the activity of Factor Xa. Both the activity offree Factor Xa and Factor Xa assembled in the prothrombinase complex(Factor Xa, Factor Va, calcium and phospholipid) are inhibited bycompounds of formula 1. The inhibition of the Factor Xa activity isobtained by direct complex formation between the inhibitor and theenzyme and is therefore independent of the plasma co-factor antithrombinIII. Effective inhibition of the Factor Xa activity is achieved byadministering the compounds either by oral administration, continuousintravenous infusion, bolus intravenous administration or any otherparenteral route such that it achieves the desired effect of preventingthe activity of Factor Xa induced formation of thrombin fromprothrombin.

Anticoagulant therapy is indicated for the treatment and prophylaxis ofa variety of thrombotic conditions of both the venous and arterialvasculature. In the arterial system, abnormal thrombus formation isprimarily associated with arteries of the coronary, cerebral andperipheral vasculature. The diseases associated with thromboticocclusion of these vessels principally include acute myocardialinfarction (AMI), unstable angina, thromboembolism, acute vessel closureassociated with thrombolytic therapy and percutaneous transluminalcoronary angioplasty (PTCA), transient ischemic attacks, stroke,intermittent claudication and bypass grafting of the coronary (CABG) orperipheral arteries. Chronic anticoagulant therapy may also bebeneficial in preventing the vessel luminal narrowing (restenosis) thatoften occurs following PTCA and CABG, and in the maintenance of vascularaccess patency in long-term hemodialysis patients. With respect to thevenous vasculature, pathologic thrombus formation frequently occurs inthe veins of the lower extremities following abdominal, knee and hipsurgery (deep vein thrombosis, DVT). DVT further predisposes the patientto a higher risk of pulmonary thromboembolism. A systemic, disseminatedintravascular coagulopathy (DIC) commonly occurs in both vascularsystems during septic shock, certain viral infections and cancer. Thiscondition is characterized by a rapid consumption of coagulation factorsand their plasma inhibitor resulting in the formation oflife-threatening thrombin throughout the microvasculature of severalorgan systems. The indications discussed above include some, but notall, of the possible clinical situations where anticoagulant therapy iswarranted. Those experienced in this field are well aware of thecircumstances requiring either acute or chronic prophylacticanticoagulant therapy.

These compounds may be used alone or in combination with otherdiagnostic, anticoagulant, antiplatelet or fibrinolytic agents. Forexample adjunctive administration of inhibitors of the activity ofFactor Xa with standard heparin, low molecular weight heparin, directthrombin inhibitors (i.e. hirudin), aspirin, fibrinogen receptorantagonists, streptokinase, urokinase and/or tissue plasminogenactivator may result in greater antithrombotic or thrombolytic efficacyor efficiency. The compounds described herein may be administered totreat thrombotic complications in a variety of animals such as primatesincluding humans. Inhibition of factor Xa is useful not only in theanticoagulant therapy of individuals having thrombotic conditions but isuseful whenever inhibition of blood coagulation is required such as toprevent coagulation of stored whole blood and to prevent coagulation inother biological samples for testing or storage. Thus, any inhibitor ofFactor Xa activity can be added to or contacted with any mediumcontaining or suspected of containing Factor Xa and in which it isdesired that blood coagulation be inhibited.

In addition to their use in anticoagulant therapy, inhibitors of FactorXa activity may find utility in the treatment or prevention of otherphysiological conditions in which the generation of thrombin has beenimplicated as playing a pathologic role. For example, thrombin has beenproposed to contribute to the morbidity and mortality of such chronicand degenerative diseases as arthritis, cancer, atherosclerosis,restenosis post coronary angioplasty and Alzheimer's disease by virtueof its ability to regulate many different cell types through specificcleavage and activation of a cell surface thrombin receptor. Inhibitionof factor Xa activity will effectively block thrombin generation andtherefore neutralize any pathologic effects of thrombin on various celltypes.

According to a further feature of the invention there is provided amethod for the treatment of a human or animal patient suffering from, orsubject to, a physiological condition which can be ameliorated by theadministration of an inhibitor of the Factor Xa activity, for exampleconditions as hereinbefore described, which comprises the administrationto the patient of a therapeutically effective amount of compound offormula I or a composition containing a compound of formula I."Effective amount" is meant to describe an amount of compound of thepresent invention effective in inhibiting the activity of Factor Xa andthus producing the desired therapeutic effect.

The present invention also includes within its scope pharmaceuticalformulations which comprise at least one of the compounds of formula Iin association with a pharmaceutically acceptable carrier or coating.

In practice compounds of the present invention may generally beadministered parenterally, intravenously, subcutaneouslyintramuscularly, colonically, nasally, intraperitoneally, rectally ororally.

The products according to the invention may be presented in formspermitting administration by the most suitable route and the inventionalso relates to pharmaceutical compositions containing at least oneproduct according to the invention which are suitable for use in humanor veterinary medicine. These compositions may be prepared according tothe customary methods, using one or more pharmaceutically acceptableadjuvants or excipients. The adjuvants comprise, inter alia, diluents,sterile aqueous media and the various non-toxic organic solvents. Thecompositions may be presented in the form of tablets, pills, granules,powders, aqueous solutions or suspensions, injectable solutions, elixirsor syrups, and can contain one or more agents chosen from the groupcomprising sweeteners, flavorings, colorings, or stabilizers in order toobtain pharmaceutically acceptable preparations.

The choice of vehicle and the content of active substance in the vehicleare generally determined in accordance with the solubility and chemicalproperties of the product, the particular mode of administration and theprovisions to be observed in pharmaceutical practice. For example,excipients such as lactose, sodium citrate, calcium carbonate, dicalciumphosphate and disintegrating agents such as starch, alginic acids andcertain complex silicates combined with lubricants such as magnesiumstearate, sodium lauryl sulfate and talc may be used for preparingtablets. To prepare a capsule, it is advantageous to use lactose andhigh molecular weight polyethylene glycols. When aqueous suspensions areused they can contain emulsifying agents or agents which facilitatesuspension. Diluents such as sucrose, ethanol, polyethylene glycol,propylene glycol, glycerol and chloroform or mixtures thereof may alsobe used.

For parenteral administration, emulsions, suspensions or solutions ofthe products according to the invention in vegetable oil, for examplesesame oil, groundnut oil or olive oil, or aqueous-organic solutionssuch as water and propylene glycol, injectable organic esters such asethyl oleate, as well as sterile aqueous solutions of thepharmaceutically acceptable salts, are used. The solutions of the saltsof the products according to the invention are especially useful foradministration by intramuscular or subcutaneous injection. The aqueoussolutions, also comprising solutions of the salts in pure distilledwater, may be used for intravenous administration with the proviso thattheir pH is suitably adjusted, that they are judiciously buffered andrendered isotonic with a sufficient quantity of glucose or sodiumchloride and that they are sterilized by heating, irradiation ormicrofiltration.

Suitable compositions containing the compounds of the invention may beprepared by conventional means. For example, compounds of the inventionmay be dissolved or suspended in a suitable carrier for use in anebulizer or a suspension or solution aerosol, or may be absorbed oradsorbed onto a suitable solid carrier for use in a dry powder inhaler.

Solid compositions for rectal administration include suppositoriesformulated in accordance with known methods and containing at least onecompound of formula I.

The percentage of active ingredient in the compositions of the inventionmay be varied, it being necessary that it should constitute a proportionsuch that a suitable dosage shall be obtained. Obviously, several unitdosage forms may be administered at about the same time. The doseemployed will be determined by the physician, and depends upon thedesired therapeutic effect, the route of administration and the durationof the treatment, and the condition of the patient. In the adult, thedoses are generally from about 0.01 to about 100, preferably about 0.01to about 10, mg/kg body weight per day by inhalation, from about 0.01 toabout 100, preferably 0.1 to 70, more especially 0.5 to 10, mg/kg bodyweight per day by oral administration, and from about 0.01 to about 50,preferably 0.01 to 10, mg/kg body weight per day by intravenousadministration. In each particular case, the doses will be determined inaccordance with the factors distinctive to the subject to be treated,such as age, weight, general state of health and other characteristicswhich can influence the efficacy of the medicinal product.

The products according to the invention may be administered asfrequently as necessary in order to obtain the desired therapeuticeffect. Some patients may respond rapidly to a higher or lower dose andmay find much weaker maintenance doses adequate. For other patients, itmay be necessary to have long-term treatments at the rate of 1 to 4doses per day, in accordance with the physiological requirements of eachparticular patient. Generally, the active product may be administeredorally 1 to 4 times per day. It goes without saying that, for otherpatients, it will be necessary to prescribe not more than one or twodoses per day.

Compounds within the scope of the present invention exhibit markedpharmacological activities according to tests described in theliterature which tests results are believed to correlate topharmacological activity in humans and other mammals. The followingpharmacological test results are typical characteristics of compounds ofthe present invention.

Enzyme Assays

The ability of the compounds in the present invention to act asinhibitors of factor Xa, thrombin, trypsin, tissue-plasminogen activator(t-PA), urokinase-plasminogen activator (u-PA), plasmin and activatedprotein C is evaluated by determining the concentration of inhibitorwhich resulted in a 50% loss in enzyme activity (IC50) using purifiedenzymes.

All enzyme assays are carried out at room temperature in 96-wellmicrotiter plates using a final enzyme concentration of 1 nM. Theconcentrations of factor Xa and thrombin are determined by active sitetitration and the concentrations of all other enzymes are based on theprotein concentration supplied by the manufacturer. Compounds accordingto the invention are dissolved in DMSO, diluted with their respectivebuffers and assayed at a maximal final DMSO concentration of 1.25%.Compound dilutions are added to wells containing buffer and enzyme andpre-equilibrated for between 5 and 30 minutes. The enzyme reactions areinitiated by the addition of substrate and the color developed from thehydrolysis of the peptide-p-nitroanilide substrates is monitoredcontinuously for 5 minutes at 405 nm on a Vmax microplate reader(Molecular Devices). Under these conditions, less than 10% of thesubstrate is utilized in all assays. The initial velocities measured areused to calculate the amount of inhibitor which resulted in a 50%reduction of the control velocity (IC50). The apparent Ki values arethen determined according to the Cheng-Prusoff equation (IC50=Ki 1+S!/Km!) assuming competitive inhibition kinetics.

By way of example, 3- 3-(S)-(Benzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate has a Ki value of 14 nM.

By way of example, 3-2-Oxo-3-(S)-(5-pyridin-4-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate has a Ki value of 55 nM.

An additional in vitro assay may be used to evaluate the potency ofcompounds according to the invention in normal human plasma. Theactivated partial thromboplastin time is a plasma-based clotting assaythat relies on the in situ generation of factor Xa, its assembly intothe prothrombinase complex and the subsequent generation of thrombin andfibrin which ultimately yields the formation of a clot as the assayendpoint. This assay is currently used clinically to monitor the ex vivoeffects of the commonly used anticoagulant drug heparin as well asdirect acting antithrombin agents undergoing clinical evaluation.Therefore, activity in this in vitro assay is considered as a surrogatemarker for in vivo anticoagulant activity.

Human Plasma Based Clotting Assay

Activated partial thromboplastin clotting times are determined induplicate on a MLA Electra 800 instrument. A volume of 100 μl ofcitrated normal human pooled plasma (George King Biomedical) is added toa cuvette containing 100 μl of a compound according to the invention inTris/NaCl buffer (pH 7.5) and placed in the instrument. Following a 3minute warming period the instrument automatically adds 100 μl ofactivated cephaloplastin reagent (Actin, Dade) followed by 100 μl of0.035M CaCl₂ to initiate the clotting reaction. Clot formation isdetermined spectrophotometrically and measured in seconds. Compoundpotency is quantitated as the concentration required to double a controlclotting time measured with human plasma in the absence of the compoundaccording to the invention.

A compound according to the invention may also be evaluated for their invivo antithrombotic efficacy in two well established animal experimentalmodels of acute vascular thrombosis. A rabbit model of jugular veinthrombosis and a rat model of carotid artery thrombosis are used todemonstrate the antithrombotic activity of these compounds in distinctanimal model paradigms of human venous thrombosis and arterialthrombosis, respectively.

Experimental In Vivo Rabbit Venous Thrombosis Model

This is a well characterized model of fibrin rich venous thrombosis thatis validated in the literature and shown to be sensitive to severalanticoagulant drugs including heparin (Antithrombotic Effect ofRecombinant Truncated Tissue Factor Pathway Inhibitor (TFPI 1-161) inExperimental Venous Thrombosis-a Comparison with Low Molecular WeightHeparin, J. Holst, B. Lindblad, D. Bergqvist, O. Nordfang, P. B.Ostergaard, J. G. L. Petersen, G. Nielsen and U. Hedner. Thrombosis andHaemostasis, 71, 214-219 (1994). The purpose of utilizing this model isto evaluate the ability of compounds to prevent the formation of venousthrombi (clots) in vive generated at a site of injury and partial stasisin the jugular vein.

Male and female New Zealand white rabbits weighing 1.5-2 kg areanesthetized with 35 mg/kg of ketamine and 5 mg/kg xylazine in a volumeof 1 ml/kg (i.m.). The right jugular vein is cannulated for infusion ofanesthetic (ketamine/xylazine 17/2.5 mg/kg/hr at a rate of approximately0.5 ml/hr) and administration of test substances. The right carotidartery is cannulated for recording arterial blood pressure andcollecting blood samples. Body temperature is maintained at 39° C. witha GAYMAR T-PUMP. The left external jugular vein is isolated and all sidebranches along an exposed 2-3 cm of vessel are tied off. The internaljugular vein is cannulated, just above the bifurcation of the commonjugular, and the tip of the cannula is advanced just proximal to thecommon jugular vein. A 1 cm segment of the vein is isolated withnon-traumatic vascular clamps and a relative stenosis is formed by tyinga ligature around the vein with an 18G needle just below the distal mostclamp. This creates a region of reduced flow and partial stasis at theinjury site. The isolated segment is gently rinsed with saline 2-3 timesvia the cannula in the internal jugular. Thereafter the isolated segmentis filled with 0.5 ml of 0.5% polyoxyethylene ether (W-1) for 5 minutes.W-1 is a detergent which disrupts the endothelial cell lining of thesegment, thus providing a thrombogenic surface for initiating clotformation. After 5 minutes the W-1 is withdrawn from the segment, andthe segment is again gently rinsed with saline 2-3 times. The vascularclamps are then removed, restoring blood flow through this portion ofthe vessel. Clot formation is allowed to form and grow for 30 minutesafter which the vein is cut just below the stenotic ligature andinspected for blood flow (the absence of blood flow is recorded ascomplete occlusion). The entire isolated segment of vein is then ligatedand the formed clot is removed and weighed (wet weight). The effect oftest agents on final clot weights is used as the primary end point.Animals are maintained for an additional thirty minutes to obtain afinal pharmacodynamic measure of anticoagulation. Drug administration isinitiated 15 minutes prior to vascular injury with W-1 and continuedthrough the period of clot formation and maturation. Three blood samples(3 ml ea.) are obtained for evaluation of hemostatic parameters: onejust prior to administration of W-1; a second 30 minutes after removalof the vascular clamps and a third at the termination of the experiment.Antithrombotic efficacy is expressed as a reduction in the final clotweight in preparations treated with a compound according to theinvention relative to vehicle treated control animals.

Experimental In Vivo Rat Arterial Thrombosis Model

The antithrombotic efficacy of factor Xa inhibitors againstplatelet-rich arterial thrombosis may be evaluated using a wellcharacterized rat carotid artery FeCl₂ -induced thrombosis model(Superior Activity of a Thromboxane Receptor Antagonist as Compared withAspirin in Rat Models of Arterial and Venous Thrombosis, W. A.Schumacher, C. L. Heran, T. E. Steinbacher, S. Youssef and M. L.Ogletree. Journal of Cardiovascular Pharmacology, 22, 526-533 (1993);Rat Model of Arterial Thrombosis Induced by Ferric Chloride, K. D.Kurtz, B. W. Main, and G. E. Sandusky. Thrombosis Research, 60, 269-280(1990); The Effect of Thrombin Inhibition in a Rat Arterial ThrombosisModel, R. J. Broersma, L. W. Kutcher and E. F. Heminger. ThrombosisResearch 64, 405-412 (1991). This model is widely used to evaluate theantithrombotic potential of a variety of agents including heparin andthe direct acting thrombin inhibitors.

Sprague Dawley rats weighing 375-450 g are anesthetized with sodiumpentobarbital (50 mg/kg i.p.). Upon reaching an acceptable level ofanesthesia, the ventral surface of the neck is shaved and prepared foraseptic surgery. Electrocardiogram electrodes are connected and lead IIis monitored throughout the experiment. The right femoral vein andartery are cannulated with PE-50 tubing for administration of a compoundaccording to the invention and for obtaining blood samples andmonitoring blood pressure, respectively. A midline incision is made inthe ventral surface of the neck. The trachea is exposed and intubatedwith PE-240 tubing to ensure airway patency. The right carotid artery isisolated and two 4-0 silk sutures are placed around the vessel tofacilitate instrumentation. An electromagnetic flow probe (0.95-1.0 mmlumen) is placed around the vessel to measure blood flow. Distal to theprobe a 4×4 mm strip of parafilm is placed under the vessel to isolateit from the surrounding muscle bed. After baseline flow measurements aremade, a 2×5 mm strip of filter paper previously saturated in 35% FeCl₂is placed on top of the vessel downstream from the probe for ten minutesand then removed. The FeCl₂ is thought to diffuse into the underlyingsegment of artery and cause deendothelialization resulting in acutethrombus formation. Following application of the FeCl₂ -soaked filterpaper, blood pressure, carotid artery blood flow and hear rate aremonitored for an observation period of 60 minutes. Following occlusionof the vessel (defined as the attainment of zero blood flow), or 60minutes after filter paper application if patency is maintained, theartery is ligated proximal and distal to the area of injury and thevessel is excised. The thrombus is removed and weighed immediately andrecorded as the primary end point of the study.

Following surgical instrumentation a control blood sample (B1) is drawn.All blood samples are collected from the arterial catheter and mixedwith sodium citrate to prevent clotting. After each blood sample, thecatheter is flushed with 0.5 ml of 0.9% saline. A compound according tothe invention is administered intravenously (i.v.) starting 5 minutesprior to FeCl₂ application. The time between FeCl₂ application and thetime at which carotid blood flow reached zero is recorded as time toocclusion (TTO). For vessels that did not occlude within 60 minutes, TTOis assigned a value of 60 minutes. Five minutes after application ofFeCl₂, a second blood sample is drawn (B2). After 10 minutes of FeCl₂exposure, the filter paper is removed from the vessel and the animal ismonitored for the remainder of the experiment. Upon reaching zero bloodflow blood a third blood sample is drawn (B3) and the clot is removedand weighed. Template bleeding time measurements are performed on theforelimb toe pads at the same time that blood samples are obtained.Coagulation profiles consisting of activated partial thromboplastin time(APTT) and prothrombin time (PT) are performed on all blood samples. Insome instances a compound according to the invention may be administeredorally. Rats are restrained manually using standard techniques andcompounds are administered by intragastric gavage using a 18 gaugecurved dosing needle (volume of 5 ml/kg). Fifteen minutes afterintragastric dosing, the animal is anesthetized and instrumented asdescribed previously. Experiments are then performed according to theprotocol described above.

The present invention may be embodied in other specific forms withoutdeparting from the spirit or essential attributes thereof.

What is claimed is:
 1. A compound of formula I ##STR19## ##STR20## isphenyl or monocyclic heteroaryl; R is hydrogen, optionally substitutedalkyl, optionally substituted aralkyl, optionally substitutedheteroaralkyl, R₆ O(CH₂)_(x) --, R₆ O₂ C(CH₂)_(x) --, Y¹ Y²NC(O)(CH₂)_(x) --, or Y¹ Y² N(CH₂)_(x) --;R₁ is hydrogen, alkyl,hydroxy, alkoxy, Y¹ Y² N--, halogen, --CO₂ R₆, --C(O)NY¹ Y², --(CH₂)_(x)OR₆, --(CH₂)_(x) NY¹ Y², or --CN; R₂ and R₃ are independently selectedfrom hydrogen, hydroxy, alkoxy, Y¹ Y² N--, halogen, --CO₂ R₆, C(O)NY¹Y², --(CH₂)_(x) OR₆, --(CH₂)_(x) NY¹ Y², --CN, optionally substitutedalkyl, optionally substituted cycloalkyl, optionally substitutedheterocyclyl, optionally substituted aryl, optionally substitutedheteroaryl, optionally substituted aralkyl, optionally substitutedheteroaralkyl, optionally substituted aralkenyl or optionallysubstituted heteroaralkenyl, or R₂ and R₃ taken together with the carbonatoms through which they are linked form an optionally substituted 5 to7 membered fused cycloalkyl, optionally substituted 5 to 7 memberedfused heterocyclyl ring or an optionally substituted 6 membered fusedaryl, or an optionally substituted 5 to 7 membered fused heteroarylring; R₄ is hydrogen or optionally substituted lower alkyl, optionallysubstituted aralkyl or optionally substituted heteroaralkyl; X₁ andX_(1a) are independently selected from hydrogen, optionally substitutedalkyl, optionally substituted aryl, optionally substituted aralkyl,optionally substituted heteroaryl or optionally substitutedheteroaralkyl, or X₁ and X_(1a) taken together form oxo; X₂ and X_(2a)are hydrogen, or taken together form oxo; X₃ is hydrogen, hydroxy,optionally substituted alkyl, optionally substituted aryl, optionallysubstituted heteroaryl, optionally substituted aralkyl or optionallysubstituted heteroaralkyl, or X₃ and one of X₁ and X_(1a) taken togetherwith the carbon atoms through which X₃ and one of X₁ and X_(1a) arelinked form a 4 to 7 membered cycloalkyl or heterocyclyl ring; X₄ ishydrogen, optionally substituted alkyl or an optionally substitutedaralkyl; X₅ and X_(5a) are hydrogen or taken together are ═NR₅ ; R₅ ishydrogen, R₆ O₂ C--, R₆ O--, cyano, R₆ CO--, optionally substitutedlower alkyl, nitro or Y¹ Y² N--; Y¹ and Y² are independently hydrogen,optionally substituted alkyl, optionally substituted aryl, optionallysubstituted aralkyl or optionally substituted heteroaralkyl, or Y¹ andY² taken together with the N through which Y¹ and Y² are linked form a 4to 7 membered heterocyclyl; X₆ and X_(6a) are independently hydrogen, R₇R₈ N--, R₉ O--, R₇ R₈ NCO--, R₇ R₈ NSO₂ --, R₇ R₈ NSO₂ N--, R₇ R₈ SO₂O--, R₉ CO--, --CO₂ R₆, --C(O)NY¹ Y², --(CH₂)_(x) CO₂ R₆, --(CH₂)_(x)C(O)NY¹ Y², --(CH₂)_(x) OR₆, --(CH₂)_(x) NY¹ Y², halo, cyano or nitro;R₆ is hydrogen, optionally substituted alkyl, optionally substitutedaralkyl or optionally substituted heteroaralkyl; R₇ and R₈ areindependently hydrogen or optionally substituted lower alkyl, or one ofR₇ and R₈ is hydrogen and the other of R₇ and R₈ is R₁₀ (O)CCH₂ -- orlower acyl; R₉ is hydrogen, optionally substituted lower alkyl,optionally substituted lower acyl or R₁₀ (O)CCH₂ --; R₁₀ is hydrogen,optionally substituted lower alkyl, optionally substituted alkoxy orhydroxy; A is S or --CH═CH--; m is 0, 1, 2 or 3; n is 0, 1, 2 or 3; andx is 1, 2, 3, 4, or 5, ora pharmaceutically acceptable salt thereof, anN-oxide thereof, a hydrate thereof or a solvate thereof.
 2. The compoundof claim 1 wherein n=1 and m=1.
 3. The compound of claim 1 wherein X₂and X_(2a) taken together are oxo.
 4. The compound of claim 1 whereinX₁, X_(1a), X₄ are hydrogen, and X₃ is hydrogen or alkyl.
 5. Thecompound of claim 1 wherein X₅ and X_(5a) taken together are ═NR₅wherein R₅ is R₆ O₂ C--.
 6. The compound of claim 1 wherein X₅ andX_(5a) taken together are ═NR₅ wherein R₅ is --OH.
 7. The compound ofclaim 1 wherein X₅ and X_(5a) taken together are ═NR₅ wherein R₅ is H.8. The compound of claim 1 wherein ##STR21## is phenyl and the carbonsubstituted with X₅, X_(5a) and R₄ HN-- is attached meta relative to theattachment of the --(CH)_(n) N-- moiety.
 9. The compound of claim 1wherein ##STR22## is thienyl and the carbon substituted with X₅, X_(5a)and R₄ HN-- is attached in the 2 position relative to the sulfur thereofand the attachment of the --(CH)_(n) N-- moiety in the 4 position. 10.The compound of claim 1 wherein R is hydrogen, methyl, aralkyl,heteroaralkyl, HO₂ CCH₂ --, H₂ NC(O)CH₂ --, or R₆ HNC(O)CH₂ --.
 11. Thecompound of claim 1 wherein R₁ is hydrogen, alkyl, or halogen.
 12. Thecompound of claim 1 wherein R₂ and R₃ are independently hydrogen,halogen, alkyloxy, amino, aryl, or heteroaryl.
 13. The compound of claim1 wherein R₂ and R₃ form an optionally substituted fused aryl or anoptionally substituted fused heteroaryl ring wherein the substituent ishalogen, alkyl, amino, hydroxy, or alkoxy.
 14. The compound of claim 1wherein R₂ and R₃ form an optionally substituted fused cycloalkyl or anoptionally substituted fused heterocyclyl in which the heteroatom isnitrogen wherein the substituent is hydrogen, Y¹ Y² N, or alkyl.
 15. Thecompound of claim 1 wherein wherein ##STR23## is phenyl and one of X₆and X_(6a) is amino or hydroxy in a para position relative to the##STR24## moiety.
 16. The compound of claim 1 wherein X₆ and X_(6a) arehydrogen.
 17. A compound according to claim 1 wherein A is S.
 18. Acompound according to claim 1 wherein A is --CH═CH-- and R₂ and R₃ forman optionally substituted 6 membered heteroaryl ring in which the heteroatom is N and the substituent is chloro, hydroxy or amino.
 19. Acompound according to claim 1 which is3- 3-(S)-(Benzob!thiophene-2-sulfonylamino)-oxo-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 3- 3-(S)- (Benzob!thiophene-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 4- 3-(S)-(Benzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-thiophene-2-carboxamidinetrifluoroacetate; 4- 3-(S)- (Benzob!thiophene-2-sulfonyl!-methylamino!-2-oxo-pyrrolidin-1-ylmethyl!-thiophene-2-carboxamidinetrifluoroacetate; 3- 3-(S)-(4-Chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 3- 3-(S)-(6-Chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 3- 3-(S)- (4-Chlorobenzob!thiophene-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 3- 3-(S)- (6-Chlorobenzob!thiophene-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 3- 3-(S)-(5-Chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 3- 3-(S)- (5-Chlorobenzob!thiophene-2-sulfonyl)-methyl-amino!-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 3- 3-(S)-(4-Methylbenzob!thiophene-2-sulfonylamino)-2-oxo -pyrrolidin -1-ylmethyl!-benzamidinetrifluoroacetate; 3- 3-(S)-(6-Methylbenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 3- 3-(S)-(5-Methylbenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 3- 3-(S)-(4,6-Dichlorobenzob!thiophene-2-sulfonylamino)-2oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; ( 3- 3-(S)-(4,6-Dichlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-phenyl!-iminomethyl)-carbamicacid 2,2,2-trichloroethyl ester; 4-Amino-3- 3-(S)-(4,6-dichlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 4Hydroxy-3- 3-(S)-(4,6-dichlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 3- 3-(S)-(6-Fluorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 4-Amino-3- 3-(S)-(6-fluorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 4-Hydroxy-3- 3-(S)-(6-fluorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 4-Amino-3- 3-(S)-(4-chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 4-Hydroxy-3- 3-(S)-(4-chlorobenzob!thiophene-2-sulfamoylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 3- 3-(S)-(4-Chloro-thieno3,2-c!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 4-Hydroxy-3- 3-(S)-(4-chloro-thieno3,2-c!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 3- 3-(S)-(5-Chlorothieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 3- 3-(S)-(Thieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinebistrifluoroacetate; 3- 3-(S)-(5-Chlorothieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-hydroxybenzamidinetrifluoroacetate; 3-{3-(S)- (5-Chlorothieno3,2-b!pyridine-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl}-benzamidinetrifluoroacetate; 3- 3-(S)-(6-Chlorothieno2,3-b!pyridine-2-sulfonylamino)-2oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 3- 3-(S)-(Thieno2,3-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinebistrifluoroacetate; 3-{3-(S)- (6-Chlorothieno2,3-b!pyridine-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1ylmethyl}-benzamidinetrifluoroacetate; 4-Hydroxy-3- 2-oxo-3-(S)-(5-chlorothieno3,2-b!pyridine-2-sulfonylamino)-pyrrolidin 1-ylmethyl}-benzamidinetrifluoroacetate; 4-Hydroxy-3- 2-oxo-3-(S)-(thieno3,2-b!pyridine-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinebistrifluoroacetate; 4-Hydroxy-3- 2-oxo-3-(S)-(5-chlorothieno3,2-b!pyridine-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-hydroxybenzamidinetrifluoroacetate; 4-Amino-3- 2-oxo-3-(S)-(5-chlorothieno3,2-b!pyridine-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 4-Amino-3- 2-oxo-3-(S)-(thieno3,2-b!pyridine-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinebistrifluoroacetate; 4- 3-(S)-(5-Chlorothieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-thiophene-2-carboxamidinetrifluoroacetate; 4- 3-(S)-(5-Chlorothieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-thiophene-2-hydroxycarboxamidinetrifluoroacetate; 4-{3-(S)- (5-Chlorothieno3,2-b!pyridine-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl}-thiophene-2-carboxamidinetrifluoroacetate; 3-{3-(S)-5-(2-Methylsulfanyl-pyrimidin-4-yl)-thiophene-2-sulfonylamino!-2-oxo-pyrrolidin-1-ylmethyl}-benzamidinetrifluoroacetate; 3-{3-(S)-5-(2-Methoxy-pyrimidin-4-yl)-thiophene-2-sulfonylamino!-2-oxo-pyrrolidin-1-ylmethyl}-benzamidinetrifluoroacetate; 3-{3-(S)-5-(2-Amino-pyrimidin-4-yl)-thiophene-2-sulfonylamino!-2-oxo-pyrrolidin-1-ylmethyl}-benzamidineditrifluoroacetate; 3-{3-(S)-(5-(2-Amino-pyrimidin-4-yl)-thiophene-2-sulfonyl!-methylamino)-2-oxo-pyrrolidin-1-ylmethyl}-benzamidineditrifluoroacetate; 3- 3-(S)-(5'-Chloro-2,2'!-bithiophenyl-5-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 4-Amino-3- 3-(S)-benzob!thiophene-2-sulfonylamino)-2-oxopyrrolidin-1-ylmethyl!benzamidinetrifluoroacetate; 4-Amino-3- 6-chlorobenzob!thiophene-2-sulfonylamino)-2oxopyrrolidin-1-ylmethyl!benzamidinetrifluoroacetate; 4-Amino-3-6-chlorobenzob!thiophene-2-sulfonylamino)-2-oxopyrrolidin-1-ylmethyl!hydroxybenzamidinetrifluoroacetate; 3-2-Oxo-3-(S)-(5-pyridin-3-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 4-Amino-3-2-oxo-3-(S)-(5-pyridin-3-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 4-Hydroxy-3-2-oxo-3-(S)-(5-pyridin-3-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 4-Hydroxy-3-2-oxo-3-(S)-(5-pyridin-N-oxide-3-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 3-2-Oxo-3-(S)-(5-pyridin-4-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 3-3-(S)-(4-Chloro-thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 3-{3-(S)-5-(5-Chloropyridin-3-yl)-thiophene-2-sulfonylamino!-2-oxopyrrolidin-1-ylmethyl}-benzamidinetrifluoroacetate; 3-3-(S)-(4-Chloro-5-pyridin-3-ylthiophene-2-sulfonylamino)-2-oxopyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 4-Hydroxy-3- 3-(S)-(6-chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidetrifluoroacetate; 3-3-(S)-(1-Aminoisoquinoline-6sulfonylamino)-2-oxopyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate; 4-Fluoro-3-3-(S)-(5-pyridin-3-yl-thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidetrifluoroacetate; 2Chloroquinoline-6-sulfonic acid {1-3-(aminoiminomethyl)-benzyl!-2-oxo-pyrrolidin-3-(S)-yl}-amidetrifluoroacetate; or 2-Aminoquinoline-6-sulfonic acid {1-3-(aminoiminomethyl)-benzyl!-2-oxo-pyrrolidin-3-(S)-yl}-amideditrifluoroacetate.
 20. A compound according to claim 19 which is3-3-(S)- (Benzob!thiophene-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.
 21. A compound according to claim 19 which is4-3-(Benzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-thiophene-2-carboxamidinetrifluoroacetate.
 22. A compound according to claim 19 which is3-3-(S)-(6-Chlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.
 23. A compound according to claim 19 which is3- 3-(S)-(6-Chlorobenzob!thiophene-2-sulfonyl)-methylamino!-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.
 24. A compound according to claim 19 which is3-3-(4,6-Dichlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.
 25. A compound according to claim 19 which is( 3-3-(S)-(4,6-Dichlorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-phenyl!-iminomethyl)-carbamicacid 2,2,2-trichloroethyl ester.
 26. A compound according to claim 19which is3- 3-(6-Fluorobenzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.
 27. A compound according to claim 19 whichis4-Amino-3- 2-oxo-3-(S)-(6-fluorobenzob!thiophene-2-sulfamoylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.
 28. A compound according to claim 19 whichis4-Hydroxy-3- 2-oxo-3-(S)-(6-fluorobenzob!thiophene-2-sulfamoylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.
 29. A compound according to claim 19 whichis4-Hydroxy-3- 2-oxo-3-(S)-(4-chlorothieno3,2-c!pyridine-2-sulfamoylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.
 30. A compound according to claim 19 which is3-3-(S)-(Thieno3,2-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidineditrifluoroacetate.
 31. A compound according to claim 19 which is3-3-(S)-(Thieno2,3-b!pyridine-2-sulfonylamino)-2-oxo-pyrrolidin-1-ylmethyl!-benzamidineditrifluoroacetate.
 32. A compound according to claim 19 whichis4-Amino-3- 2-oxo-3-(S)-(thieno3,2-b!pyridine-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidineditrifluoroacetate.
 33. A compound according to claim 19 whichis3-{3-(S)-5-(2-Methylsulfanyl-pyrimidin-4-yl)-thiophene-2-sulfonylamino!-2-oxo-pyrrolidin-1-ylmethyl}-benzamidinetrifluoroacetate.
 34. A compound according to claim 19 whichis4-Amino-3- 6-Chlorobenzob!thiophene-2-sulfonylamino)-2-oxopyrrolidin-1-ylmethyl!benzamidinetrifluoroacetate.
 35. A compound according to claim 19 whichis4-Amino-3- 6-Chlorobenzob!thiophene-2-sulfonylamino)-2-oxopyrrolidin-1-ylmethyl!hydroxybenzamidinetrifluoroacetate.
 36. A compound according to claim 19 which is3-2-Oxo-3-(S)-(5-pyridin-3-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.
 37. A compound according to claim 19 whichis4-Hydroxy-3-2-oxo-3-(S)-(5pyridin-3-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.
 38. A compound according to claim 19 whichis4Hydroxy-3-2-oxo-3-(S)-(5pyridin-N-oxide-3-yl-thiophene-2-sulfonylamino)-pyrrolidin-1-ylmethyl!-benzamidinetrifluoroacetate.
 39. A compound according to claim 19 whichis4Hydroxy-3- 3-(S)-(6-chloro-benzob!thiophene-2-sulfonylamino)-2-oxo-pyrrolidin-1-yl!benzamidetrifluoroacetate.
 40. A pharmaceutical composition comprising atherapeutically effective amount of the compound according to claim 1and a pharmaceutically acceptable carrier.
 41. A method for treating apatient suffering from a physiological disorder capable of beingmodulated by inhibiting an activity of Factor Xa comprisingadministering to the patient a therapeutically effective amount of thecompound according to claim
 1. 42. The method according to claim 41wherein the physiological disorder is venous vasculature, arterialvasculature, abnormal thrombus formation, acute myocardial infarction,unstable angina, thromboembolism, acute vessel closure associated withthrombolytic therapy, percutaneous transluminal coronary angioplasty,transient ischemic attacks, stroke, intermittent claudication or bypassgrafting of the coronary or peripheral arteries, vessel luminalnarrowing, restenosis post coronary or venous angioplasty, maintenanceof vascular access patency in long-term hemodialysis patients,pathologic thrombus formation occurring in the veins of the lowerextremities following abdominal, knee and hip surgery, a risk ofpulmonary thromboembolism, or disseminated systemic intravascularcoagulopathy occurring in vascular systems during septic shock, certainviral infections or cancer.
 43. The method according to claim 41 whereinthe physiological disorder abnormal thrombus formation, acute myocardialinfarction, unstable angina, thromboembolism, acute vessel closureassociated with thrombolytic therapy, transient ischemic attacks,intermittent claudication or bypass grafting of the coronary orperipheral arteries, restenosis post coronary or venous angioplasty,pathologic thrombus formation occurring in the veins of the lowerextremities following abdominal, knee and hip surgery or a risk ofpulmonary thromboembolism.
 44. The method according to claim 41 whereinthe physiological disorder stroke, vessel luminal narrowing, maintenanceof vascular access patency in long-term hemodialysis patients, ordisseminated systemic intravascular coagulopathy occurring in vascularsystems during septic shock, certain viral infections or cancer.